Anti-EIF2B5 (phospho S539) antibody

• WB

Supplier Data

Western blot - Anti-EIF2B5 (phospho S539) antibody (AB4775)

A 80 kDa band corresponding to Phospho-eIF2b epsilon was observed across the cell lines tested and reduced upon treatment and in differentiated cell line.

All lanes:

Western blot - Anti-EIF2B5 (phospho S539) antibody (ab4775) at 1/2000 dilution

Lane 1:

L6 whole cell extract at 30 µg

Lane 2:

L6 differentiated to myotubes at 30 µg

Lane 3:

L6 treated with Insulin (10nM for 10 min) at 30 µg

Lane 4:

C2C12 at 30 µg

Lane 5:

C2C12 treated with Insulin (10nM for 10 min) at 30 µg

Secondary

All lanes:

Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/4000 dilution

Predicted band size: 80 kDa

false

• WB

Unknown

Western blot - Anti-EIF2B5 (phospho S539) antibody (AB4775)

Peptide Competition : Recombinant eIF-2B episilon treated with lambda phosphatase (1) or treated with GSK-3 beta (lanes 2-5) and added to background extracts were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4^oC, then were incubated with 0.50 μg/mL ab4775 antibody, following prior incubation with : no peptide (1, 2), the nonphosphopeptide corresponding to the immunogen (3), a generic phophoserine containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and signals were detected using the Tropix WesternStar method. The data show that only the peptide corresponding to ab4775 blocks the antibody signal, thereby demonstrating the specificity of the antibody. The data also show the up-regulation of the ab4775 phospho signal upon stimulation with GSK-3 beta. Pep

All lanes:

Western blot - Anti-EIF2B5 (phospho S539) antibody (ab4775)

Predicted band size: 80 kDa

false