Synthetic peptide derived from the region of human RPS6 that contains serines 244 and 247 (unfortunately, the amino acid sequence is considered to be commercially sensitive)
HeLa +/- TNF-alpha or anisomycin
Centrifuge before opening to settle contents.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Preservative: 0.05% Sodium Azide
Constituents: Dulbecco's PBS without magnesium and calcium, 50% Glycerol, PBS, 1mg/ml BSA, pH 7.3
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Immunogen affinity purified
Purified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated RPS6. The final product is generated by
affinity chromatography using a RPS6-derived peptide that is phosphorylated at serines 244 and 247.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Detects a band of approximately 32 kDa (predicted molecular weight: 31 kDa).
May play an important role in controlling cell growth and proliferation through the selective translation of particular classes of mRNA.
Belongs to the ribosomal protein S6e family.
Ribosomal protein S6 is the major substrate of protein kinases in eukaryote ribosomes. The phosphorylation is stimulated by growth factors, tumor promoting agents, and mitogens. It is dephosphorylated at growth arrest.
Lysates prepared from HeLa cells left untreated (1) or treated with TNF-alpha (2-5) were resolved by SDS-PAGE on a 14% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer for one hour at room temperature, and incubated with ab12865 for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 2), the non-phosphopeptide corresponding to the immunogen (3), a generic phosphoserine-containing peptide (4), or the phosphopeptide immunogen (5). After washing, membranes were incubated with goat anti-rabbit IgG HRP conjugate and bands were detected. The data show that only the peptide corresponding to RPS6 [phospho S244 & phospho S247] blocks the signal, verifying the specificity of the antibody.