Cd49d 9f10

Manufactured by BD

CD49d (9F10) is a monoclonal antibody that binds to the alpha-4 subunit (CD49d) of the VLA-4 (Very Late Antigen-4) integrin. VLA-4 is involved in cell-cell and cell-extracellular matrix interactions.

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Lab products found in correlation

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Close spelling variants of this product

Anti-CD49d (clone 9F10, CD49d (clone 9F10) APC-conjugated mouse anti-human CD49d (clone 9F10, CD49d-FITC (clone 9F10, CD49d BV421 (9F10,

4 protocols using cd49d 9f10

Flow Cytometry Antibody Staining Protocol

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The following antibodies were used for culturing or surface and intracellular cytokine staining (ICS) for flow cytometry: CD28 (CD28.2, BD), CD49d (9F10, BD), CD3-PB (SP34-2, BD), CD4-BV510 (L200, BD), CD8-PB (RPA-T8, BD), IFN-γ-APC (4S.B3, BD), IFN-γ Brilliant Violet 711 (4S.B3, Biolegend), TNF-α-PE (MAb11, BD), TNF-α-PE-Cy7 (MAb11, BD), IL-17-PE (eBio64CAP17, eBioscience), IL-22-biotinylated (anti-human IL-22, RD), Streptavidin-Pacific blue (Invitrogen), Perforin-biotinylated (Pf-344, Mabtech), Caspase 3-AF647 (C92-605,BD), and anti-Vγ2-FITC (7A5, Pierce).
After staining, the cells were fixed and subjected to flow cytometry. Lymphocytes were gated based on forward- and side-scatters, and at least 40,000 gated events were analyzed using Summit Data Acquisition and Analysis Software (DakoCytomation).

Yang E., Yang R., Guo M., Huang D., Wang W., Zhang Z., Chen C., Wang F., Ho W., Shen L., Xiao H., Chen Z.W, & Shen H. (2018). Multidrug-resistant tuberculosis (MDR-TB) strain infection in macaques results in high bacilli burdens in airways, driving broad innate/adaptive immune responses. Emerging Microbes & Infections, 7, 207.

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Multiparameter Flow Cytometry Panels

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The following Abs were used for culture or surface and intracellular cytokine staining for flow cytometry: CD28 (CD28.2, BD), CD49d (9F10, BD), CD3-PECY7 (SP34-2, BD), CD3-PB (SP34-2, BD), CD4-APC (L200, BD, used for measuring CD4 expression after CD4 depleting Ab treatment), CD8-PB (RPA-T8, BD), CD8-APC (RPA-T8, BD), CD8-PE (RPA-T8, BD), IFN-γ-Allophycocyanin (4S.B3, BD), IFN-γ-PE (4S.B3, BD), TNF-α-PE (MAb11, BD), TNF-α-APC (MAb11, BD), TNF-α-PB (MAb11, eBioscience), IL-17-PE (eBio64CAP17, eBioscience), IL-17-Alexa647 (eBio64CAP17, eBioscience), IL-22-biotinylated (anti-human IL-22, RD, see reference(44 (link))), Streptavidin-Pacific blue (invitrogen). IL-4-APC (8D4-8, eBioscience), Perforin-biotinylated (Pf-344, Mabtech)

Yao S., Huang D., Chen C.Y., Halliday L., Wang R.C, & Chen Z.W. (2014). CD4+ T cells are required to contain early extrathoracic TB dissemination and sustain multi-effector functions of CD8+ T and CD3− lymphocytes. Journal of immunology (Baltimore, Md. : 1950), 192(5), 2120-2132.

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Phenotyping Antigen-Specific CD8+ T Cells

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PBMCs were isolated from fresh whole blood samples by Ficoll density centrifugation. 1x10⁶ PBMCs were stimulated with 1 μg of peptide in 1 mL of RPMI 1640 supplemented with 10% FBS and 10U IL-2 in a 48-well plate for six days. 10U of IL-2 were supplemented every other day. After six days, cells were stained with KK10 or SL9 pentamer and re-stimulated overnight in 1 mL of RPMI 1640 supplemented with 10% FBS, 10U IL-2, 0.5 μg CD28 (CD28.2, BD Biosciences), 0.5 μg CD49d (9F10, BD Biosciences), 3 μM monensin, 1 μg brefeldin A, and 1 μg of peptide. After stimulation for 15 hours, PBMCs were stained with CD3 (UCHT1, Biolegend), CD8 (RPA-T8, Biolegend), IFN-γ (B27, Biolegend), perforin (B-D48, Cell Sciences) and KK10 or SL9 pentamer (Proimmune) before analysis by flow cytometry (BDFACS Canto II) and analysis (FlowJo, TreeStar). Due to availability, samples were run in single replicates.

Pohlmeyer C.W., Laskey S.B., Beck S.E., Xu D.C., Capoferri A.A., Garliss C.C., May M.E., Livingston A., Lichmira W., Moore R.D., Leffell M.S., Butler N.J., Thorne J.E., Flynn J.A., Siliciano R.F, & Blankson J.N. (2018). Cross-reactive microbial peptides can modulate HIV-specific CD8+ T cell responses. PLoS ONE, 13(2), e0192098.

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Cytokine Analysis by Flow Cytometry

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The following Abs were used for culture or surface and intracellular cytokine staining for flow cytometry: CD28 (CD28.2, BD), CD49d (9F10, BD), CD3-Pacific blue (SP34-2,BD), CD4-BV510 (L200, BD), CD8- Pacific blue (RPA-T8, BD), IFN-γ-APC (4S.B3, BD), IFN-γ Brilliant Violet 711 (4S.B3, Biolegend), TNF-α-PE (Mab11, BD), TNF-α-PE-Cy7 (Mab11, BD), IL-17-PE (eBio64CAP17, eBioscience), IL-22-biotinylated (anti-human IL-22, RD), Streptavidin-Pacific blue (invitrogen), Perforin-biotinylated (Pf-344, Mabtech), Caspase 3-AF647 (C92-605,BD), anti-Vγ2-FITC (7A5, Pierce).
After staining, cells were fixed and subjected to analysis on flow cytometer of BD LSRFortessa^TM Cell Analyzer. Lymphocytes were gated based on forward- and side-scatters, and at least 40,000 gated events were analyzed using Summit Data Acquisition and Analysis Software (Dako Cytomation).

Shen H., Yang E., Guo M., Yang R., Huang G., Peng Y., Sha W., Wang F, & Shen L. (2022). Adjunctive Zoledronate + IL-2 administrations enhance anti-tuberculosis Vγ2Vδ2 T-effector populations, and improve treatment outcome of multidrug-resistant tuberculosis1. Emerging Microbes & Infections, 11(1), 1790-1805.

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