{"product_id":"mouse-cd4-t-cell-isolation-kit-column-free-s0k3003","title":"Mouse CD4 T Cell Isolation Kit (Column-Free)","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\u003e1 mL Mouse CD4 T Cell Biotin-Antibody Cocktail\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e0.75 mL Streptavidin Beads\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\u003cp\u003e\u003cspan\u003eSuitable for a total cell count of 1×10⁹, and can perform up to 100 sorting runs.\u003c\/span\u003e\u003c\/p\u003e\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\u003eIn this negative selection process, non- is labeled with a biotin-conjugated antibody.\u003c\/span\u003e\u003cspan style=\"color:#404040\"\u003eCD4 T\u003c\/span\u003e\u003cspan\u003eCells are then incubated with streptavidin‑coated nanomagnetic beads; non‑target cells are labeled via antibody complexes bound to the beads, while the target…\u003c\/span\u003e\u003cspan style=\"color:#404040\"\u003eCD4 T\u003c\/span\u003e\u003cspan\u003eThe cells remain unlabeled. The cell suspension is then transferred to a polystyrene tube and placed within the sorter's magnetic field. Under the influence of the magnetic field, the magnetically labeled non-target cells adhere to the tube wall, while the unlabeled\u003c\/span\u003e\u003cspan style=\"color:#404040\"\u003eCD4 T\u003c\/span\u003e\u003cspan\u003eThe cells remain in the supernatant. This supernatant is, therefore, the solution from which non-target cells have been removed.\u003c\/span\u003e\u003cspan style=\"color:#404040\"\u003eCD4 T\u003c\/span\u003e\u003cspan\u003eCell-enriched fraction.\u003c\/span\u003e\u003c\/p\u003e\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%;\"\u003e150 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-Free\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%;\"\u003eNegative 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; do not freeze.\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 style=\"text-align:left\"\u003e\u003cspan style=\"color:#000000\"\u003eMouse CD4 T lymphocytes are a core functional cell subset of the murine adaptive immune system. They originate from hematopoietic stem cells in the bone marrow, complete their development and maturation in the thymus, and then migrate to secondary lymphoid organs such as the spleen and lymph nodes. These cells specifically express the CD4 molecule on their surface and can recognize antigen peptides presented by antigen‑presenting cells (APCs) in complex with MHC class II molecules via the T‑cell receptor (TCR), thereby becoming activated and exerting immunoregulatory functions; they are often referred to as the \"command cells\" of the immune system. In scientific research, obtaining highly purified, high‑activity mouse CD4 T cells is essential for conducting related studies. This negative selection kit employs a negative selection strategy: biotin‑conjugated antibodies label non‑CD4⁺ T cells, which are then bound by streptavidin‑coated magnetic nanoparticles and removed, keeping CD4 T cells untouched throughout the process. This approach maximizes the preservation of their native morphology and functional activity, providing reliable cellular material for subsequent experiments.\u003c\/span\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cspan style=\"display:inline; white-space:nowrap;\"\u003eNote: This kit does not include sorting buffer; please prepare it yourself. The corresponding Stater product number is\u003ca href=\"https:\/\/www.starter-bio.com\/s0d3018_starter_magsep_separation_buffer.html\" style=\"color:#0066FF; display:inline; white-space:nowrap;\"\u003eS0D3018\u003c\/a\u003e\u003c\/span\u003e\u003ctable style=\"border-collapse:collapse;width:auto;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;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 #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eOperating Instructions\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eDosage and Time\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 #000000;border:1px solid #000000;border:1px solid #000000;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 #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eCollect the mouse spleen, grind it on a 40 μm sterile filter to prepare a single-cell suspension of splenic cells; perform 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; splenic cells can be centrifuged at 400 g for 7 minutes;\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003ePrepare the sample\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 #000000;border:1px solid #000000;border:1px solid #000000;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 #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eResuspend 1×10 in 100μL MagSep Separation Buffer\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e7\u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003ecells;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote:\u003c\/span\u003e\u003cspan\u003eAdjust reagents proportionally according to the sample volume; set aside a pre-sorting sample to measure the initial CD4 ratio;\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e1×10\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e7\u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003ecells\/100 μL\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 #000000;border:1px solid #000000;border:1px solid #000000;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 #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eAdd 10 μL Mouse CD4 T cell Biotin Antibody Cocktail to the sample and gently mix.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote:\u003c\/span\u003e\u003cspan\u003eIn this step, gentle pipetting 2–3 times can be used to mix evenly.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e10 μL \/ 100 μL\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 #000000;border:1px solid #000000;border:1px solid #000000;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 #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eIncubate the antibodies in the cocktail with the cells at room temperature for 10 min;\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eIncubate at room temperature for 10 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 #000000;border:1px solid #000000;border:1px solid #000000;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 #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eShake for 5–30 s to mix the Streptavidin beads thoroughly;\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eMix for 5–30 s\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 #000000;border:1px solid #000000;border:1px solid #000000;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 #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eAdd 7.5 μL of Streptavidin beads to the sample;\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e7.5 μL\/100 μL\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 #000000;border:1px solid #000000;border:1px solid #000000;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 #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eGently mix the magnetic beads and cells, and incubate at room temperature for 5 minutes.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote:\u003c\/span\u003e\u003cspan\u003eIn this step, you can use a pipette to gently blow the mixture 2–3 times for mixing.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eDuring this step, the magnetic beads may settle at the bottom; you can gently tap the tube to remix them after about 2.5 minutes.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eIncubate at room temperature for 5 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 #000000;border:1px solid #000000;border:1px solid #000000;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 #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eAdd MagSep Separation Buffer to the sample and immediately place it on the magnetic stand to prevent the magnetic beads from settling heavily at the bottom.\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eBring the total volume up to 2.5 mL (for a 5 mL flow cytometry tube).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eAdd the total volume to 7.5 mL (using a 15 mL centrifuge tube)\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;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 #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003ePlace the sample on the magnetic stand so that the magnetic beads adhere to the tube wall;\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eAdsorb using a single-well magnetic stand (3 min)\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eAdsorb using a multi-well magnetic stand (5 min)\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;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 #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eTilt the magnetic stand and pour the sample into a new collection tube to collect the sample;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eNote: The sorted cells can be centrifuged at 400g for 7 minutes, for subsequent culture and analysis.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eCell sorting successful\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\u003ctd colspan=\"3\" style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote: 1.\u003c\/span\u003e\u003cspan\u003eFor 0.1–1 × 10^8 cells (0.1–1 mL), it is recommended to use a 5 mL single-well magnetic separator (Starter EasyIso Separator) with a total adsorption volume of 2.5 mL.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eFor 2–5 × 10^8 cells (2–5 mL), it is recommended to use a 15 mL multi-well magnetic separator (Starter EasyEights EasyIso Separator) with a total adsorption volume of 7.5 mL.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e2.\u003c\/span\u003e\u003cspan\u003eIf the live cell rate in the sample is low, it is recommended to use a dead cell removal kit to eliminate dead cells (\u003c\/span\u003e\u003cspan style=\"text-decoration:underline;color:#000000\"\u003eDead Cell Removal Set, (RUO)\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e#S0K0009\u003c\/span\u003e\u003cspan\u003e);\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\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":"100T","offer_id":43030516662347,"sku":"S0K3003-100T","price":0.1,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/78c303c07f444929863c341b3fbdb902.jpg?v=1786510892","url":"https:\/\/www.antbioinc.com\/products\/mouse-cd4-t-cell-isolation-kit-column-free-s0k3003","provider":"AntBio","version":"1.0","type":"link"}