Anti-JAK1 (phospho Y1034 + Y1035) antibody (ab5493)

Western blot - JAK1 (phospho Y1034 + Y1035) antibody (ab5493)

Peptide Competition:
Extracts prepared from 3T3L1 cells left unstimulated (1) or stimulated with LIF (2-5) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then incubated with ab5493 antibody for two hours at room temperature in a 3% BSATBST buffer, following prior incubation with: no peptide (1, 2), the non-phosphopeptide corresponding to the immunogen (3), a generic phosphotyrosine-containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignal method. The data show that the peptide corresponding to ab5493 blocks the antibody signal, thereby demonstrating the specificity of the antibody. The data also show the induction of phosphorylation by the addition of LIF.

Extracts prepared from 3T3L1 cells left unstimulated (1) or stimulated with LIF (2-5) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then incubated with ab5493 antibody for two hours at room temperature in a 3% BSATBST buffer, following prior incubation with: no peptide (1, 2), the non-phosphopeptide corresponding to the immunogen (3), a generic phosphotyrosine-containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignal method. The data show that the peptide corresponding to ab5493 blocks the antibody signal, thereby demonstrating the specificity of the antibody. The data also show the induction of phosphorylation by the addition of LIF.