Anti-EIF2S1 (phospho S51) antibody (ab4837)

Western blot - EIF2S1 (phospho S51) antibody (ab4837)

Peptide Competition:

Extracts prepared from 3T3-L1 adipocytes 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 4oC, then were incubated with 0.50 µg/mL ab4837 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with:
1) the phosphopeptide immunogen
2) the non-phosphopeptide corresponding to the immunogen
3) no peptide

After washing, membranes were incubated with goat F(ab)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStar method. The data show that only the peptide corresponding to ab4837 blocks the antibody signal, thereby demonstrating the specificity of the antibody.

Peptide Competition:

Extracts prepared from 3T3-L1 adipocytes 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 4oC, then were incubated with 0.50 µg/mL ab4837 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with:
1) the phosphopeptide immunogen
2) the non-phosphopeptide corresponding to the immunogen
3) no peptide

After washing, membranes were incubated with goat F(ab)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStar method. The data show that only the peptide corresponding to ab4837 blocks the antibody signal, thereby demonstrating the specificity of the antibody.

Immunohistochemistry (Frozen sections) - EIF2S1 (phospho S51) antibody (ab4837)

ab4837 staining eIF2S1(phospho S51) in mouse skeletal muscle tissue sections (5 µm) by IHC-Fr (Frozen sections). Tissue samples were fixed with acetone (for 10 minutes at -20°C) and blocked with 2% BSA for 1 hour at 25°C. The sample was incubated with primary antibody (1/250 in PBS with 1% BSA) at 4°C for 9 hours. An Alexa Fluor^®488-conjugated Goat polyclonal to rabbit IgG (1/500) was used as secondary antibody.

This image is courtesy of an anonymous Abreview

Immunocytochemistry/ Immunofluorescence - EIF2S1 (phospho S51) antibody (ab4837)

ab4837 staining eIF2S1 in human 293ft cells by Immunocytochemistry/ Immunofluorescence. The cells were paraformaldehyde fixed, permeabilised in 0.1% Triton X-100 and then blocked using 5% serum for 1 hour at 25°C. Samples were then incubated with primary antibody at 1/200 for 2 hours at 25°C. The secondary antibody used was a goat anti-rabbit IgG conjugated to Alexa Fluor® 488 (green) used at a 1/250 dilution. DAPI was used to stain the cell nuclei (blue).

This image is courtesy of an anonymous abreview.