Peptide Competition and Phosphatase Stripping:
Extracts of NIH3T3 cells stably expressing c-Src Y527F mutant protein were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was left untreated (1-4) or treated with lambda (λ) phosphatase (5), blocked with a 5% BSA-TBST buffer overnight at 4°C, then incubated with the cortactin [pY466] (mouse) antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 5), the non-phosphopeptide corresponding to the phosphopeptide immunogen (2), a generic phosphotyrosine-containing peptide (3), or the phosphopeptide immunogen (4). After washing, the membrane was incubated with goat F(ab')2 anti-rabbit IgG alkaline phosphatase (cat.# ALI4405) and signals were detected using the Tropix WesternStar™ method.
Human peripheral blood leukocytes stained with CD45 FITC and antibody clone 581 PerCP-Cy®5.5 conjugate (top, Cat. No. A14949) or mouse IgG1,kappa PerCP-Cy®5.5 isotype control (bottom). Cy®togram gated for total live CD14 negative lymphocyte population. Go ›
Human peripheral blood leukocytes stained with CD34 antibody (clone 581) Pacific Blue conjugate (top, Cat. No. A14947) or mouse IgG1,kappa Pacific Blue isotype control (bottom) and costained with CD45 FITC, CD14 PE, and 7-AAD, then analyzed via gating on total live and CD14-negative lymphocyte population. Go ›