Post Translational Modification
Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
Anti-CREB pS133 antibody reacts with phosphorylated human CREB at pS133 and shows minimal reactivity by western blot with non-phosphorylated CREB and minimal reactivity (1%) by ELISA against the non-phosphorylated form of the immunizing peptide. This antibody was assayed against a variety of tissues including fibroblasts and B-cell lysates. Bands of 46 and 43 kDa corresponding to phosphorylated CREB are observed in western blots. Anti-CREB pS133 is suitable for the detection by immunoblot of phosphoryl-ated human, mouse and rat CREB. No cross-reactivity occurs with non-phosphorylated CREB. For immunohistochemistry, formalin fixed, paraffin embedded human tissue shows moderate to strong nuclear staining in a variety of cells with minimal background staining. Although not tested this antibody is likely functional by FACS and immunoprecipitation.
Anti-CREB pS133 was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first sequentially pre-adsorbed against a control E.coli lysate, nickel-purified recombinant CREB and the non-phosphorylated form of the immunizing peptide. The resultant depleted antiserum was then purified against the phosphorylated form of the immunizing peptide. This phospho specific polyclonal antibody is specific for phosphorylated pS133 of human CREB. Reactivity with non-phosphorylated CREB is minimal. This antibody does show cross reactivity with pS133 phosphorylated CREB from mouse and rat.
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