BD Pharmingen- PE Hamster Anti-Mouse CD69_BD Pharmingen
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Flow cytometric analysis of CD69 expression on stimulated mouse splenocytes. Mouse splenic leucocytes were stimulated for 5 hours at 37°C with Phorbol 12-Myristate 13-Acetate (PMA, 10 ng/mL; Sigma-Aldrich, Cat. No. P-8139). The stimulated cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with either PE Hamster IgG1, λ1 Isotype Control (Cat. No. 553954; dashed line histogram) or PE Hamster Anti-Mouse CD69 antibody (Cat. No. 561932/553237; solid line histogram) at 0.06 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing CD69 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) stimulated leucocytes. Flow cytometry and data analysis was performed using a BD FACSCelesta™ Flow Cytometer System and FloJo™ software.
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