![Lysates from Jurkat cells either untreated (1, 6), treated with hydrogen peroxide (2-5), or ionophore A23187 (7) were resolved on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 3% milk-TBST buffer for one hour at room temperature, then were incubated with the ETS1[pS 282] antibody overnight at 4%deg;C in a 1% milk-TBST buffer, following prior incubation with: no peptide (1-2, 6-7), the non-phosphopeptide corresponding to the immunogen (3), a generic phosphoserine-containing peptide (4), or the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab′)2 anti-rabbit IgG HRP conjugate (Cat. ##ALI4404), and bands were detected using the Pierce SuperSignal™ method.](http://www.bioprodhub.com/system/product_images/ab_products/55/sub_4/7083_441109Gimg01-20150203155824.jpg)
Lysates from Jurkat cells either untreated (1, 6), treated with hydrogen peroxide (2-5), or ionophore A23187 (7) were resolved on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 3% milk-TBST buffer for one hour at room temperature, then were incubated with the ETS1[pS 282] antibody overnight at 4%deg;C in a 1% milk-TBST buffer, following prior incubation with: no peptide (1-2, 6-7), the non-phosphopeptide corresponding to the immunogen (3), a generic phosphoserine-containing peptide (4), or the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab′)2 anti-rabbit IgG HRP conjugate (Cat. ##ALI4404), and bands were detected using the Pierce SuperSignal™ method.