![Left panel: Extracts of Jurkat cells untreated (1) or treated with 10 µM hydrogen peroxide for 10 minutes (2-5) were resolved on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 3% milk-TBST buffer for one hour at room temperature, then incubated with the ETS1[pT38] antibody overnight at 4°C in a 1% milk-TBST buffer, following prior incubation with: no peptide (1, 2), the non-phosphopeptide corresponding to the phosphopeptide immunogen (3), a generic phosphothreonine-containing peptide (4), or the phosphopeptide immunogen (5). After washing, the membrane was incubated with goat F(ab') 2 anti-rabbit IgG HRP conjugate (Cat. ##ALI4404) and signals were detected using the Pierce SuperSignal™ method. The data show that only the phosphopeptide corresponding to ETS1[pT38] blocks the signal, verifying the specificity of the antibody. The data also show up-regulation of the signal upon hydrogen peroxide treatment in this cell system. Right panels: Extracts of Jurk](http://www.bioprodhub.com/system/product_images/ab_products/55/sub_4/7955_441104Gimg01-20150203155824.jpg)
Left panel: Extracts of Jurkat cells untreated (1) or treated with 10 µM hydrogen peroxide for 10 minutes (2-5) were resolved on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 3% milk-TBST buffer for one hour at room temperature, then incubated with the ETS1[pT38] antibody overnight at 4°C in a 1% milk-TBST buffer, following prior incubation with: no peptide (1, 2), the non-phosphopeptide corresponding to the phosphopeptide immunogen (3), a generic phosphothreonine-containing peptide (4), or the phosphopeptide immunogen (5). After washing, the membrane was incubated with goat F(ab') 2 anti-rabbit IgG HRP conjugate (Cat. ##ALI4404) and signals were detected using the Pierce SuperSignal™ method. The data show that only the phosphopeptide corresponding to ETS1[pT38] blocks the signal, verifying the specificity of the antibody. The data also show up-regulation of the signal upon hydrogen peroxide treatment in this cell system. Right panels: Extracts of Jurk