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Anti c kit clone 2b8

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Anti-c-Kit (clone 2B8) is a monoclonal antibody that binds to the c-Kit receptor, also known as CD117. It is used for the detection and analysis of c-Kit-expressing cells in flow cytometry and other immunoassays.

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Lab products found in correlation

Anti sca1 clone d7, by Thermo Fisher Scientific (3 mentions) Anti cd34 clone ram34, by Thermo Fisher Scientific (2 mentions) Anti cd127 clone a7r34, by Thermo Fisher Scientific (2 mentions) Anti flk 2 clone a2f10, by Thermo Fisher Scientific (2 mentions) Anti fcεriα clone mar 1, by BioLegend (1 mentions) Anti cd8a clone 53 6, by Thermo Fisher Scientific (1 mentions) Dynal beads, by Thermo Fisher Scientific (1 mentions) Histopaque 1083, by Merck Group (1 mentions) Facsaria 3, by BD (1 mentions) Anti cd16 cd32 clone 2.4g2, by BD (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Flow cytometry, by Thermo Fisher Scientific (1 mentions) Streptavidin pe cy7, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

APC-conjugated anti-c-Kit (clone 2B8; Anti-mouse CD117 (c-kit) APC (clone 2B8), C-Kit (clone 2B8; C-Kit (2B8; Anti-c-kit Kit c

4 protocols using anti c kit clone 2b8

1

Generation of Bone Marrow-Derived Mast Cells

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Bone marrow-derived cultured mast cells (BMCMCs) were generated from bone marrow cells of female C57BL/6J mice by culture in Wehi-3-conditioned, IL-3-containing medium (DMEM containing 20% supernatant of Wehi-3 cells, 10% FCS, 50 μM β-mercaptoethanol, 2 mM L-glutamine, and 1% antibiotic-antimycotic solution) as previously described (Kalesnikoff and Galli, 2011 (link)). The cells were used after at least 4 weeks of culture. Flow cytometry was used to confirm c-Kit and FcεRIα expression (~95 % of cells were c-Kit- and FcεRIα-positive) using anti-c-Kit (clone 2B8: eBioscience) and anti-FcεRIα (clone Mar-1: Biolegend) antibodies; these cultures thus predominately consisted of mast cells.
Ho C.C., Chhabra A., Starkl P., Schnorr P.J., Wilmes S., Moraga I., Kwon H.S., Gaudenzio N., Sibilano R., Wehrman T.S., Gakovic M., Sockolosky J.T., Tiffany M.R., Ring A.M., Piehler J., Weissman I.L., Galli S.J., Shizuru J.A, & Garcia K.C. (2017). Decoupling the Functional Pleiotropy of Stem Cell Factor by Tuning c-Kit Signaling. Cell, 168(6), 1041-1052.e18.
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2

Isolation and Characterization of Murine Hematopoietic Stem Cells

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Mononuclear BM cells were isolated by low-density centrifugation (Histopaque 1083, Sigma) and stained with a cocktail of biotinylated lineage antibodies. Biotinylated antibodies used for lineage staining were all rat anti-mouse antibodies: anti-CD11b (clone M1/70), anti-B220 (clone RA3-6B2), anti-CD5 (clone 53-7.3), anti-Gr-1 (clone RB6-8C5), anti-Ter119 and anti-CD8a (Clone 53-6.7) (all from eBioscience). After lineage depletion by magnetic separation (Dynalbeads, Invitrogen), cells were stained with anti-Sca-1 (clone D7) (eBioscience), anti-c-kit (clone 2B8) (eBioscience), anti-CD34 (clone RAM34) (eBioscience), anti-CD127 (clone A7R34) (eBioscience), anti-Flk-2 (clone A2F10) (eBioscience), and Streptavidin (eBioscience). Early haematopoiesis FACS analysis data were plotted as percentage of long-term haematopoietic stem cells (LT-HSCs, gated as LSK CD34−/lowFlk2−), short-term haematopoietic stem cells (ST-HSCs, gated as LSK CD34+Flk2−), and lymphoid-primed multipotent progenitors (LMPPs, gated as LSK CD34+Flk2+) distribution among LSKs (Linnegc-kit+sca-1+ cells). In order to isolate HSCs, lineage depletion was performed to enrich for lineage negative cells. Lineage negative cells were then stained as aforementioned and sorted using a BD FACS Aria III (BD Bioscience). More information on the antibodies used is provided in the Source Data file.
Montserrat-Vazquez S., Ali N.J., Matteini F., Lozano J., Zhaowei T., Mejia-Ramirez E., Marka G., Vollmer A., Soller K., Sacma M., Sakk V., Mularoni L., Mallm J.P., Plass M., Zheng Y., Geiger H, & Florian M.C. (2022). Transplanting rejuvenated blood stem cells extends lifespan of aged immunocompromised mice. NPJ Regenerative Medicine, 7, 78.
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3

Hematopoietic Stem Cell Characterization

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Freshly isolated stroma‐enriched cells were co‐culture with BM lineage‐negative depleted cell at the ratio 1:1 in growth factor‐free medium IMDM 10% FBS at 37°C, 5% CO2, 3% O2 for 72 h. After 24 h in culture, BrdU was added to the cells at a final concentration of 1 mM. At the endpoint, cells were harvested by intensive washing with HBSS and incubated with Cell dissociation buffer enzyme‐free based (Gibco, Life Technology) for 30–50 min at 37°C. After being counted, cells were stained with the HSC surface markers: anti‐Sca‐1 (clone D7) (eBioscience), anti‐c‐Kit (clone 2B8) (eBioscience), anti‐CD34 (clone RAM34) (eBioscience), anti‐CD127 (clone A7R34) (eBioscience), anti‐CD16/CD32 (clone 2.4G2) (BD Biosciences), anti‐Flk‐2 (clone A2F10) (eBioscience) and streptavidin (eBioscience). After washing, cells were stained with anti‐Annexin V antibody (BD Horizon) to identify apopotic cells rate. Finally after fixation and permeabilization, the cells were stained with antibody anti‐BrdU to verify the cell cycle status. The analysis was performed on a LSRII flow cytometer (BD Biosciences).
Guidi N., Sacma M., Ständker L., Soller K., Marka G., Eiwen K., Weiss J.M., Kirchhoff F., Weil T., Cancelas J.A., Florian M.C, & Geiger H. (2017). Osteopontin attenuates aging‐associated phenotypes of hematopoietic stem cells. The EMBO Journal, 36(7), 840-853.
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4

Quantifying Mobilized Stem/Progenitor Cells

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C57BL/6 male mice were treated with potential CXCR4 antagonists individually by subcutaneous injection, and then blood samples containing mobilized stem/progenitor cells were collected 2 h later. After labeling with specific antibodies, including APC-conjugated anti-CXCR4 (clone 2B11; eBioscience), FITC-conjugated anti-CD34 (clone RAM34; eBioscience), PE-conjugated anti-CD133 (clone 13A4; eBioscience), and anti-KDR (clone Avsa12a1; eBioscience), anti-c-Kit (clone 2B8; eBioscience), anti-Sca-1 (clone D7; eBio-science), anti-lineage (mouse hematopoietic lineage biotin panel, eBioscience), and Streptavidin PE-Cy7 (eBioscience), cells were washed, characterized, and quantified by flow cytometry (Guava Technologies, Hayward, CA). Each data point included at least 60 000 events for analysis of mobilized cells.
Wu C.H., Wang C.J., Chang C.P., Cheng Y.C., Song J.S., Jan J.J., Chou M.C., Ke Y.Y., Ma J., Wong Y.C., Hsieh T.C., Tien Y.C., Gullen E.A., Lo C.F., Cheng C.Y., Liu Y.W., Sadani A.A., Tsai C.H., Hsieh H.P., Tsou L.K, & Shia K.S. (2015). Function-Oriented Development of CXCR4 Antagonists as Selective Human Immunodeficiency Virus (HIV)-1 Entry Inhibitors. Journal of medicinal chemistry, 58(3), 1452-1465.
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