Peptide Competition. Lysates prepared from HEK293 cells left untreated (1) or treated with EGF (2-6) 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 ribosomal protein S6 [pS236] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 2), the phosphopeptide corresponding to ribosomal protein S6 [pSpS244/247] (3), the non-phosphopeptide corresponding to the immunogen (4), a generic phosphoserine-containing peptide (5), or, the phosphopeptide immunogen (6). After washing, membranes were incubated with goat F(ab and quote;)2 anti-rabbit IgG HRP conjugate (Cat. no. ALI4404) and bands were detected using the Pierce SuperSignal™ method. The data show that only the peptide corresponding to ribosomal protein S6 [pS236] blocks the signal, verifying the specificity of the antibody.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from theregion of human RPS6 that contains serine 236. The sequence is conserved in mouse and rat.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Ribosomal protein 6 (RPS6) is a component of the 40S ribosomal subunit belonging to S6E family of ribosomal proteins. RPS6 is a key substrate for kinases, and is phosphorylated by growth factors and mitogens during in cell growth and cell division. Phosporylation in RPS6 is well regulated, and the different phosphorylation sites are highly conserved. Major phosphorylation sites of RPS6 include Ser 235, 236, 240, and 244.
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