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2 protocols using anti mouse cd62l clone mel 14

1

Multiparametric Flow Cytometry of CD8+ T Cells

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FC receptors were blocked with anti-mouse CD16/32 (clone 93, Biolegend) before staining. Dead cells were identified using the Zombie Aqua Fixable Viability Kit (Biolegend). Cells were stained with the following antibody clones: anti-mouse CD8a (clone 53-6.7, Biolegend), anti-mouse/human CD44 (clone IM7, Biolegend), anti-mouse CD62L (clone MEL-14, eBioscience), anti-mouse/human Granzyme B (clone GB11, Biolegend), anti-mouse CTLA4 (clone UC1-4B9, Biolegend), anti-mouse CD25 (clone PC61.5, eBioscience), and anti-mouse IFN-γ (clone XMG1.2, Biolegend). Intracellular staining of Granzyme B, CTLA-4, and IFN-γ was performed using the Foxp3/Transcription Factor Staining Buffer Set (eBioscience). To count cells, 123count eBeads (eBioscience) were added to flow cytometry tubes immediately before flow cytometer acquisition. Data was acquired on a BD LSRFortessa and analyzed in FlowJo. Cells were gated for size, single cells, living cells, and CD8+ cells before examination of proliferation curves (Supplementary Fig. 8c). Cells were further gated on proliferation dye+ cells to exclude any CD8+ T cells in the APC fraction before quantification of division numbers and examination of surface and intracellular proteins. Statistical analyses of results from separate mice were performed using GraphPad Prism software.
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2

Multiparametric Flow Cytometry of CD8+ T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
FC receptors were blocked with anti-mouse CD16/32 (clone 93, Biolegend) before staining. Dead cells were identified using the Zombie Aqua Fixable Viability Kit (Biolegend). Cells were stained with the following antibody clones: anti-mouse CD8a (clone 53-6.7, Biolegend), anti-mouse/human CD44 (clone IM7, Biolegend), anti-mouse CD62L (clone MEL-14, eBioscience), anti-mouse/human Granzyme B (clone GB11, Biolegend), anti-mouse CTLA4 (clone UC1-4B9, Biolegend), anti-mouse CD25 (clone PC61.5, eBioscience), and anti-mouse IFN-γ (clone XMG1.2, Biolegend). Intracellular staining of Granzyme B, CTLA-4, and IFN-γ was performed using the Foxp3/Transcription Factor Staining Buffer Set (eBioscience). To count cells, 123count eBeads (eBioscience) were added to flow cytometry tubes immediately before flow cytometer acquisition. Data was acquired on a BD LSRFortessa and analyzed in FlowJo. Cells were gated for size, single cells, living cells, and CD8+ cells before examination of proliferation curves (Supplementary Fig. 8c). Cells were further gated on proliferation dye+ cells to exclude any CD8+ T cells in the APC fraction before quantification of division numbers and examination of surface and intracellular proteins. Statistical analyses of results from separate mice were performed using GraphPad Prism software.
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