Cd11b biotin

Manufactured by Thermo Fisher Scientific

Sourced in United States

CD11b-biotin is a cell surface marker that binds to the CD11b antigen. It is commonly used in flow cytometry applications to identify and enumerate myeloid cells, such as monocytes and granulocytes, within a heterogeneous cell population.

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Anti-CD11b-biotin Biotin-conjugated anti-CD11b antibody CD11b

4 protocols using cd11b biotin

Murine Model of Experimental Autoimmune Encephalomyelitis

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Myelin oligodendrocyte glycoprotein 35-55 (MOG_35-55) peptide (MEVGWYRSPFSRVVHLYRNGK) was generated by the Protein Core laboratory of the Blood Research Institute, BloodCenter of Wisconsin. The 2.4G2 hybridoma was obtained from the American Tissue Culture Collection. Anti-mouse CD4-APC-eFluor 780, CD25-Alexa Fluor 700, CD11b-PE, CD11b-biotin, IL-17-Alexa Fluor 647, CD11c-Biotin, B220-Biotin, CD8-Biotin, CD11b-biotin, Foxp3-PE and streptavidin-PE Cy5.5 were purchased from eBioscience (San Diego, CA). Anti-mouse B220-PE-Texas Red, IFN-γ-PE, anti-active caspase 3-FITC and anti-human Ki67-FITC were purchased from BD Biosciences (San Diego, CA). Anti-mouse Ly-6C-APC and Ly-6G-APC-Cy7 were purchased from Biolegend (San Diego, CA). Anti-mouse/rat Neuropilin-1-APC was obtained from R&D Systems (Minneapolis, MN). Monoclonal antibodies SMI-32 (anti-nonphosphorylated neurofilament-H) and SMI-99 (anti-myelin basic protein (MBP)) were purchased from Covance (Emeryville, CA). Strepavidin Alexa 405 and goat anti-mouse Alexa 546 (H + L) were purchased from Life Sciences Advanced Technologies (St. Petersburg, FL). Anti-Biotin microbeads were purchased from MiltenyiBiotec (Auburn, CA).

Ray A., Basu S., Miller N.M., Chan A.M, & Dittel B.N. (2014). An Increase in Tolerogenic Dendritic Cell and Natural T Regulatory Cell Numbers During EAE in Rras−/− Mice Results in Attenuated Disease. Journal of immunology (Baltimore, Md. : 1950), 192(11), 5109-5117.

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Characterizing Autoimmune Neuroinflammation

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Myelin oligodendrocyte glycoprotein 35–55 (MOG_35–55) peptide (MEVGWYRSPFSRVVHLYRNGK) and myelin basic protein (MBP) Ac_1–11 peptide (Ac-ASQKRPSQRSK) were generated by the Protein Core Laboratory of the Blood Research Institute, BloodCenter of Wisconsin. KYC was synthesized by Biomatik (Wilmington, DE). The 2.4G2 hybridoma was obtained from the American Tissue Culture Collection. Anti-mouse CD4-APC-eFluor 780 CD11b-PE, CD11b-biotin, IL-17-Alexa Fluor 647, Foxp3-PE and streptavidin-PE Cy5.5 were purchased from eBioscience (San Diego, CA). Anti-mouse IFN-γ-PE was purchased from BD Biosciences (San Diego, CA). Anti-mouse Ly-6C-APC and Ly-6G-APC-Cy7 were purchased from Biolegend (San Diego, CA). The SMI-32 antibody, which detects nonphosphorylated neurofilament-H was purchased from Covance (Emeryville, CA). Streptavidin Alexa 405 and goat anti-mouse Alex 456 (H + L) were purchased from Life Sciences Advanced Technologies (St. Petersburg, FL). Anti-MPO heavy chain was purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX). Anti-rabbit IgG (H + L)-HRP was purchased from Jackson ImmunoResearch (West Grove, PA). Anti-β-actin-HRP was purchased from Sigma-Aldrich (St. Louis, MO). For all experiments, the mice were age (6–8 wk) and gender matched with both sexes being utilized.

Zhang H., Ray A., Miller N.M., Hartwig D., Pritchard KA J.r, & Dittel B.N. (2015). Inhibition of Myeloperoxidase at the Peak of Experimental Autoimmune Encephalomyelitis Restores Blood-Brain-Barrier Integrity and Ameliorates Disease Severity. Journal of neurochemistry, 136(4), 826-836.

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Transfer of T cells from Zdhhc2 knockout mice

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Lymph nodes from CD45.2⁺ C57BL/6 and CD45.2⁺ Zdhhc2^−/− mice were respectively grounded and filtered through a 70 μm cell strainer into PBS to obtain cell suspensions. The negative selection immunomagnetic cell separation method was used to purify T cells by incubating with antibody mixture including CD11b-biotin (13-0112-85, eBioscience), CD19-biotin (13-0193-85, eBioscience), CD11c-biotin (13-0114-85, eBioscience), and Dynabeads™ M-280 Streptavidin (11206D, Invitrogen). Then the purified CD45.2⁺ C57BL/6 or CD45.2⁺ Zdhhc2^−/− T cells were counted, and 2 million cells were transferred intravenously into the CD45.1⁺ CD3ϵ^−/− mice, respectively. After 7 days, the transferred mice were treated with imiquimod for continuous 7 days. Then the lesioned skin were collected for flow cytometry analysis.

Zhou B., Yang W., Li W., He L., Lu L., Zhang L., Liu Z., Wang Y., Chao T., Huang R., Gu Y., Jia T., Liu Q., Tian S., Pierre P., Maeda T., Liang Y, & Kong E. (2021). Zdhhc2 Is Essential for Plasmacytoid Dendritic Cells Mediated Inflammatory Response in Psoriasis. Frontiers in Immunology, 11, 607442.

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Flow Cytometry Antibody Staining Protocol

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For the flow cytometry analyses, cells were stained with monoclonal antibodies against the following anti-mouse molecules: CD3-biotin, Ter119-biotin, GR-1-biotin, B220-biotin, CD11b-biotin, NK1.1-biotin, CD3-biotin, CD4-biotin, CD8-biotin, cKit-BV650, Sca1-PE, CD135-PerCP, CD34-APC, CD25-PE, CD44-APC, CD3-APC, CD8-PerCP, CD-BV650, CD45.1/Ly5.1-PE-Cy7, CD45.2/Ly5.2-APC-Cy7 (all from eBiosciences, CA, USA). For secondary detection of biotinylated antibodies, streptavidin conjugated with FITC or PE-Cy7 was used (eBiosciences, San Diego, CA, USA). All flow cytometry measurements (Canto II BD biosciences or LSRII BS Biosciences) were calibrated first with BD™ CompBead Plus, κ/Negative Control (BSA) Compensation Plus (7.5 µm). Recommended flow cytometry settings are specified in [17 ]. Flow cytometry analyses were performed using FlowJo software (Treestar, Ashland, OR, USA).

de Roo J.J., Chhatta A., Garcia-Perez L., Naber B.A., Vloemans S.A., Salvatori D.C., Pike-Overzet K., Mikkers H, & Staal F.J. (2022). Axin2/Conductin Is Required for Normal Haematopoiesis and T Lymphopoiesis. Cells, 11(17), 2679.

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