Anti-eIF4G1 (phospho S1108) antibody (ab4772)

Western blot - eIF4G1 (phospho S1108) antibody (ab4772)

Peptide Competition: Extracts prepared from Hek293 cells were resolved on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either treated with lambda phosphatase (5) or untreated (1-4, 6), were blocked with a 3% non-fat milk-TBST buffer overnight at 4^oC, then were incubated with 1.0 µg/mL ab4772 antibody for two hours at room temperature in a 3% non-fat milk-TBST buffer, following its prior incubation with the phosphopeptide immunogen (1), no peptide (2, 5, 6), the non phosphopeptide corresponding to the immunogen (3), or, a generic phosphoserine containing peptide (4). After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG alkaline phosphatase and the signal was detected using the Tropix WesternStar method. The data show that only the phosphopeptide corresponding to ab4772 blocks the antibody signal, demonstrating the specificity of the antibody. The data also show that phosphatase treatment eliminates the signal, verifying that the antibody is phosphospecific.

Peptide Competition: Extracts prepared from Hek293 cells were resolved on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either treated with lambda phosphatase (5) or untreated (1-4, 6), were blocked with a 3% non-fat milk-TBST buffer overnight at 4oC, then were incubated with 1.0 µg/mL ab4772 antibody for two hours at room temperature in a 3% non-fat milk-TBST buffer, following its prior incubation with the phosphopeptide immunogen (1), no peptide (2, 5, 6), the non phosphopeptide corresponding to the immunogen (3), or, a generic phosphoserine containing peptide (4). After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG alkaline phosphatase and the signal was detected using the Tropix WesternStar method. The data show that only the phosphopeptide corresponding to ab4772 blocks the antibody signal, demonstrating the specificity of the antibody. The data also show that phosphatase treatment eliminates the signal, verifying that the antibody is phosphospecific.