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Peptide Competition:
Extracts of PC12 cells unstimulated (1) or stimulated with 0.5 M sorbitol for 5 minutes (2-6) were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer overnight at 4°C, and then incubated with the ERK1&2 [pTpY 185/187] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following its prior incubation with: no peptide (1, 2), the non-phosphopeptide corresponding to the phosphopeptide immunogen (3), a generic phosphothreonine-containing peptide (4), a generic phosphotyrosine-containing peptide (5), or the phosphopeptide immunogen (6). 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.
![Extracts of PC12 cells unstimulated (1) or stimulated with 0.5 M sorbitol for 5 minutes (2-6) were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer overnight at 4°C, and then incubated with the ERK1&2 [pTpY 185/187] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following its prior incubation with: no peptide (1, 2), the non-phosphopeptide corresponding to the phosphopeptide immunogen (3), a generic phosphothreonine-containing peptide (4), a generic phosphotyrosine-containing peptide (5), or the phosphopeptide immunogen (6). 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.](http://tools.thermofisher.com/content/sfs/gallery/low/3838.jpg)
