
Western blot analysis of A431 cells treated with pervanadate (1 mM) for 30 minutes (30 ug/lane). Blots were probed with anti-IkappaBalpha (lane 1), anti-phospho-IkappaBalpha?(Tyr-42) (ab24783; lanes 2-5), or anti-phospho-IkappaBalpha (Tyr-305) (ab24784; lanes 6-9). Both anti-phospho-IkappaBalpha (Tyr-42) and anti-phospho-IkappaBalpha (Tyr-305) were used in the presence of no blocking peptide (lane 2 & 6), phospho-IkappaBalpha (Tyr-42) peptide (lane 3 & 8), phospho-IkappaBalpha (Tyr-305) peptide (lane 4 & 7), or BSA conjugated to phospho-tyrosine (lane 5 & 9). Peptides and BSA-pTyr were used at 1 ug/ml.