{"product_id":"cd25-nanobeads-kit-mouse-ruo-s0k0033","title":"CD25 Nanobeads Kit, mouse (RUO)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eFormat\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e(1)\u003c\/span\u003e\u003cspan\u003eDextran-coated iron oxide nanomagnetic beads coupled with anti-PE monoclonal antibodies are stored in a PBS buffer system containing BSA, Poloxamer 188, and NaN.\u003c\/span\u003e\u003csub\u003e\u003cspan\u003e3\u003c\/span\u003e\u003c\/sub\u003e\u003cspan\u003eIn the PBS buffer system.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"margin-right: 8px;\"\u003e(2)\u003c\/span\u003e\u003cspan\u003ePhycoerythrin (PE)-labeled mouse anti-CD25 monoclonal antibody is stored in a PBS buffer system containing BSA and Proclin300.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eCapacity\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\n\u003cp\u003e\u003cspan\u003e50T (0.5 mL magnetic beads \u0026amp; 0.1 mL antibody): Suitable for 5 × 10\u003csup\u003e\u003cspan\u003e8\u003c\/span\u003e\u003c\/sup\u003e\u003c\/span\u003e\u003cspan\u003eTotal cell count: up to 50 sorting runs can be performed.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e100T (1 mL magnetic beads \u0026amp; 0.2 mL antibody): Suitable for 1 × 10\u003csup\u003e\u003cspan\u003e9\u003c\/span\u003e\u003c\/sup\u003e\u003c\/span\u003e\u003cspan\u003eTotal cell count: up to 100 sorting runs can be performed.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003ePrinciple of Separation\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003e\u003cspan\u003eFirst, the cells are stained with a mouse anti-CD25 antibody labeled with phycoerythrin (PE), and then the PE‑CD25‑labeled cells are magnetically labeled using Anti‑PE Nanobeads. After labeling is complete, the cell suspension is loaded into a separation column placed within the sorter's magnetic field. The magnetically labeled CD25‑positive cells are retained in the column, while the unlabeled cells flow through the column, constituting the negative fraction. Once the column is removed from the magnetic field, the CD25‑positive cells that were captured by the magnetic beads are eluted, yielding the positive fraction.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eApplications in cell sorting\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eEnrichment or depletion of CD25-positive cells in mice.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eReactivity\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eMouse\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eMagnetic Bead Size\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e20 nm\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eSeparation Method\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eColumn-Based\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eSelection Strategy\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003ePositive Selection\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003eStore protected from light at 2–8°C; avoid freezing.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\u003cp\u003e\u003cspan\u003eCD25, also known as the interleukin-2 receptor alpha chain (IL-2Rα), is expressed in the early stages of thymic T cell development (CD4\u003csup\u003e\u003cspan\u003e-\u003c\/span\u003e\u003c\/sup\u003e\u003c\/span\u003e\u003cspan\u003eCD8\u003csup\u003e\u003cspan\u003e-\u003c\/span\u003e\u003c\/sup\u003e\u003c\/span\u003e\u003cspan\u003e); peripheral resting conventional T cells exhibit low CD25 expression, while CD25 is upregulated upon activation of T and B cells, with relatively low expression on activated monocytes. When present alone, CD25 has low affinity for IL-2; however, when it forms a trimeric complex—IL-2Rαβγ—with the β chain (CD122) and the γ chain (CD132)—it creates a high-affinity IL-2 receptor complex that mediates IL-2 signaling.\u003c\/span\u003e\u003cspan style=\"color:#0F1115\"\u003eThe CD4\u003csup\u003e\u003cspan\u003e+\u003c\/span\u003e\u003c\/sup\u003eCD25\u003csup\u003e\u003cspan\u003e+\u003c\/span\u003e\u003c\/sup\u003e cell population includes Foxp3\u003csup\u003e\u003cspan\u003e+\u003c\/span\u003e\u003c\/sup\u003e regulatory T cells (Treg).\u003c\/span\u003e\u003cspan\u003e \u003c\/span\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e\u003cspan style=\"font-weight:bold\"\u003eNote\u003c\/span\u003e\u003cspan\u003e: This kit does not include the sorting buffer; it must be provided separately. The corresponding Stater product number is S0D3018.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eTaking S\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eSeparation Column\u003c\/span\u003e\u003cspan\u003eas an example\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable border=\"1\" style=\"border-collapse:collapse;width:auto;border:1px solid #000000\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eSteps\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eOperating Procedure\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eDosage \/ Time\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"10\" style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eMagnetic Labeling\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e1\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eGrind a small tissue sample containing CD25-positive cells on a 40 μm sterile filter to prepare a single-cell suspension. After red blood cell lysis, perform live cell counting.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote:\u003c\/span\u003e\u003cspan\u003eIt is recommended to count cells after staining with AO\/PI.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e—\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e2\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eCentrifuge the cell suspension at 400×g for 7 minutes, and completely discard the supernatant.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e400×g, 7 min\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e3\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eResuspend every 1 × 10⁷ cells in 98 μL of buffer.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote:\u003c\/span\u003e\u003cspan\u003eThe volume of buffer should be adjusted proportionally according to the sample volume; (e.g., resuspend 2 × 10⁷ cells in 196 μL of sorting buffer).\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eResuspend the cells\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e4\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eAdd 2 μL of PE-CD25 antibody to label the cells, and mix the cell suspension thoroughly, avoiding excessive bubble formation during mixing.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote:\u003c\/span\u003e\u003cspan\u003eWhen the cell number increases, proportionally increase the volume of all reagents accordingly.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eAdd 2 μL of PE-CD25 antibody.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e5\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eMix thoroughly and incubate in the dark at 2–8°C for 10 minutes.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eIncubate for 10 minutes.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e6\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eWash each 1 × 10⁷ cells with 1 mL of buffer, centrifuge at 400 × g for 7 minutes, and completely discard the supernatant.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eAdd 1 mL of buffer to 1 × 10⁷ cells.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e7\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eResuspend each 1 × 10⁷ cells in 90 μL of buffer.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote:\u003c\/span\u003e\u003cspan\u003eAdjust the volume of the buffer proportionally according to the sample volume; (e.g., resuspend 2 × 10⁷ cells in 180 μL of sorting buffer).\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eResuspend the cells.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e8\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eAdd 10 μL of Anti-PE Nanobeads per 1 × 10⁷ cells.\u003c\/span\u003e\u003cspan style=\"color:#404040\"\u003e,\u003c\/span\u003e\u003cspan\u003eAnd mix the cell suspension thoroughly, avoiding excessive bubble formation during mixing.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote:\u003c\/span\u003e\u003cspan\u003eWhen the number of cells increases, proportionally increase the volume of all reagents (e.g., for 2 × 10⁷ cells, add 20 μL of magnetic beads).\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eFor 1 × 10⁷ cells, add 10 μL.\u003c\/span\u003e\u003cspan style=\"color:#404040\"\u003eAnti-PE Nanobeads\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e9\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eMix thoroughly and incubate in the dark at 2–8°C for 15 minutes.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eIncubate for 15 minutes.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e10\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eResuspend the cells in 500 μL of sorting buffer.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote:\u003c\/span\u003e\u003cspan\u003e500 μL of buffer can resuspend up to 1 × 10⁸ cells.\u003c\/span\u003e\u003cspan\u003eCells\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eResuspend the cells.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"5\" style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eMagnetic separation.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e11\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003ePlace the S-shaped separation column into the corresponding magnetic separator's magnetic field, and rinse the column with 500 μL of buffer.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eRinse the separation column.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e12\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eLoad the cell suspension onto the separation column and collect the flow-through (unlabeled cells).\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e—\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e13\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eWash the column with buffer three times, 500 μL each time, and add the next wash only after the liquid in the column has completely drained.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e3 × 500 μL buffer\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e14\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eCombine all eluates into the negative fraction; remove the separation column from the magnetic field and place it over a collection tube.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e—\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e15\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eAdd 1 mL of buffer to the column and quickly and firmly push down with the provided plunger to elute the magnetically labeled positive cells.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote:\u003c\/span\u003e\u003cspan\u003eFor higher purity, you can use a new separation column to repeat the magnetic separation once on the eluted positive cells.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eCollect the target cells.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eIn L\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eSeparation Column\u003c\/span\u003e\u003cspan\u003efor example\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable border=\"1\" style=\"border-collapse:collapse;width:auto;border:1px solid #000000\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eSteps\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eOperating method\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eDosage \/ Time\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"10\" style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eMagnetic labeling\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e1\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eGrind a small tissue sample containing CD25-positive cells through a 40 μm sterile filter to prepare a single-cell suspension. After red blood cell lysis, perform live cell counting.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote:\u003c\/span\u003e\u003cspan\u003eIt is recommended to count cells after staining with AO\/PI.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e—\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e2\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eCentrifuge the cell suspension at 400×g for 7 minutes, and completely aspirate the supernatant.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e400×g, 7 min\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e3\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eResuspend every 1×10⁷ cells in 98 μL of buffer.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote:\u003c\/span\u003e\u003cspan\u003eAdjust the volume of the buffer proportionally according to the sample volume; (e.g., resuspend 2×10⁷ cells in 196 μL of sorting buffer).\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eResuspend the cells.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e4\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eAdd 2 μL of PE-CD25 antibody to label the cells, mix the cell suspension thoroughly, avoiding excessive bubble formation during mixing.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote:\u003c\/span\u003e\u003cspan\u003eWhen the cell number increases, increase the volumes of all reagents proportionally.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eAdd 2 μL of PE-CD25 antibody.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e5\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eMix thoroughly and incubate in the dark at 2–8°C for 10 minutes.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eIncubate for 10 minutes.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e6\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eWash every 1×10⁷ cells with 1 mL of buffer, centrifuge at 400×g for 7 minutes, and completely aspirate the supernatant.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e1 × 10⁷ cells plus 1 mL of buffer\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e7\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eResuspend every 1 × 10⁷ cells in 90 μL of buffer\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote:\u003c\/span\u003e\u003cspan\u003eAdjust the volume of buffer proportionally according to the sample volume (e.g., resuspend 2 × 10⁷ cells in 180 μL of sorting buffer)\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eResuspend the cells\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e8\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eAdd 10 μL of Anti-PE Nanobeads per 1 × 10⁷ cells\u003c\/span\u003e\u003cspan style=\"color:#404040\"\u003e,\u003c\/span\u003e\u003cspan\u003eand mix the cell suspension; avoid generating excessive bubbles during mixing\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote:\u003c\/span\u003e\u003cspan\u003eWhen the number of cells increases, increase the volumes of all reagents proportionally (e.g., add 20 μL of magnetic beads for 2 × 10⁷ cells)\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e1 × 10⁷ cells plus 10 μL\u003c\/span\u003e\u003cspan style=\"color:#404040\"\u003eAnti-PE Nanobeads\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e9\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eMix thoroughly and incubate in the dark at 2–8°C for 15 minutes\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eIncubate for 15 minutes\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e10\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eResuspend the cells in 1 mL of sorting buffer\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote:\u003c\/span\u003e\u003cspan\u003eResuspend up to 1×10⁸ cells in 500 μL buffer\u003c\/span\u003e\u003cspan\u003eCells\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eResuspend cells\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"5\" style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eMagnetic separation\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e12\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003ePlace the L-shaped separation column into the magnetic field of the corresponding magnetic separator, and rinse the column with 3 mL of buffer\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eRinse the separation column\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e12\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eLoad the cell suspension onto the separation column and collect the flow-through (unlabeled cells)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e—\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e13\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eWash the column with buffer three times, 3 mL each time; add the next wash only after the liquid in the column has completely drained\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e3 × 3 mL buffer\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e14\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eCombine all flow-through fractions as the negative fraction; remove the separation column from the magnetic field and place it over a collection tube\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e—\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e15\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eAdd 5 mL of buffer to the column, and use the provided plunger to quickly and firmly push down to elute the magnetically labeled positive cells\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eNote: For higher purity, you can repeat the magnetic separation once on the eluted positive cells using a new separation column\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000;border:1px solid #CCCCCC;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eCollect the target cells\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Starter","offers":[{"title":"50T","offer_id":67435086151755,"sku":"S0K0033-50T","price":500.0,"currency_code":"USD","in_stock":true},{"title":"100T","offer_id":67435086184523,"sku":"S0K0033-100T","price":835.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/cd09dcb667f9483f9af0f60b4f5f83d0_9edc885a-fc0c-4e11-b28c-19755d462e86.jpg?v=1794250809","url":"https:\/\/www.antbioinc.com\/products\/cd25-nanobeads-kit-mouse-ruo-s0k0033","provider":"AntBio","version":"1.0","type":"link"}