{"product_id":"human-b-cell-isolation-kit-column-based-s0k1013","title":"Human B Cell Isolation Kit (Column-Based)","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 Human B Cell Biotin-Antibody Cocktail\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e1 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\u003eThis kit employs a column-based negative selection method to isolate B cells from human peripheral blood mononuclear cells (PBMCs). In this negative-selection process, non-B cells are labeled with biotin-conjugated antibodies, and the cells are then incubated with streptavidin-nanomagnetic beads. As a result, the non-target cells become bound to the magnetic beads via the antibody‑antigen complex, while the target B cells remain unlabeled. The cell suspension is subsequently loaded onto a separation column; under the influence of a magnetic field, the magnetically labeled non-target cells are retained within the column, whereas the unlabeled B cells elute by gravity into a collection tube. The resulting supernatant constitutes the B-cell‑enriched fraction after removal of the non-target cells.\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%;\"\u003eHuman\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%;\"\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\u003e\u003cspan style=\"color:#000000\"\u003eHuman B lymphocytes are the core effectors of adaptive humoral immune responses, performing critical functions such as antigen recognition, processing and presentation, antibody secretion, and maintenance of immunological memory. They play an irreplaceable role in defending against extracellular pathogen infections and regulating the magnitude of immune responses. The CD19 molecule is the most representative lineage-specific marker on the B-cell surface; it belongs to the immunoglobulin superfamily and is stably expressed from the early pre-B cell stage through to mature plasma cells. CD19 collaborates with the B-cell receptor to regulate B-cell activation, proliferation, and differentiation, serving as a crucial regulatory molecule for B-cell development and functional maintenance. Consequently, it is widely used as a specific surface antigen of human B cells. In immunological research, obtaining B cells that are highly pure, highly active, and have intact, native surface molecules is an essential prerequisite for ensuring the reliability of experimental results. This negative-selection kit is based on the principle of negative sorting, employing a biotin‑labeled antibody panel to specifically label non‑B cells. The identified targets include various interfering cell populations, such as T cells, NK cells, monocytes, macrophages, platelets, and erythroid cells. Streptavidin‑coated nanomagnetic beads are then used to efficiently remove non‑target cells. Throughout the entire sorting process, no antibodies bind to any antigens on the B‑cell surface, thereby maximizing the preservation of the cells' original morphology, the integrity of their surface markers, and their biological functional activity, providing high‑quality cellular material for downstream applications such as gene expression analysis, in vitro culture and expansion, and functional characterization.\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 the separation buffer; it must be provided separately. The corresponding Stater catalog 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\u003cp\u003e\u003cspan\u003eTaking\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eL Separation Column\u003c\/span\u003e\u003cspan\u003eas an example\u003c\/span\u003e\u003c\/p\u003e\n\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\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\"\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 rowspan=\"7\" 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\"\u003eCell processing and labeling\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\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\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\u003eIsolate PBMCs from peripheral blood using density gradient centrifugation with lymphocyte separation solution, or directly thaw cryopreserved PBMC cells, and then 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; PBMCs 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 1×10^7 cells in 100 μL of 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 reagent volumes proportionally based on sample volume; set aside a pre-sorting sample to determine the proportion of human B cells.\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 of Human B 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, gently pipette the mixture 2–3 times to mix thoroughly;\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\u003eMix well\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eStreptavidin Beads\u003c\/span\u003e\u003cspan\u003e, add 10 μL\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eStreptavidin Beads\u003c\/span\u003e\u003cspan\u003einto 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\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\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\u003eGently mix the magnetic beads and cells, and incubate at room temperature for 5 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 5 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\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\u003cp style=\"text-align:center\"\u003e\u003cspan\u003ePlace it in the column selection magnet,\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eL Separation Column\u003c\/span\u003e\u003cspan\u003eadd 3 mL of MagSep Separation Buffer to rinse the separation column;\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\u003eRinse the separation column\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"2\" 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\"\u003eCell sorting\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\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\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eAdd the cells to the separation column and collect the target cells using a collection tube;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote:\u003c\/span\u003e\u003cspan\u003eThe cell volume should be at least 500 μL; if insufficient, add MagSep Separation Buffer to make up the volume. The cells that drip down under gravity are the target cells.\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\u003eCollect the target 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 #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\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eAdd 3 mL of MagSep Separation Buffer and collect the target cells.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eNote:\u003c\/span\u003e\u003cspan\u003eThis step elutes the unlabeled target cells from the separation column, increasing the yield.\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\u003eCollect the target cells\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\u003ctd colspan=\"4\" 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\"\u003eNotes: 1.\u003c\/span\u003e\u003cspan\u003ePay attention to the maximum capacity of the separation column.\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eThe S Separation Column can hold a maximum of\u003c\/span\u003e\u003cspan\u003e1×10\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e7\u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003e;\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003ethe L Separation Column's maximum labeled cell count is\u003c\/span\u003e\u003cspan\u003e1×10\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e8\u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003e;\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 sample viability 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":43030515974219,"sku":"S0K1013-100T","price":0.1,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/56a866ae170a41b38b2486e016305ce3.png?v=1786510853","url":"https:\/\/www.antbioinc.com\/products\/human-b-cell-isolation-kit-column-based-s0k1013","provider":"AntBio","version":"1.0","type":"link"}