Description: Rabbit polyclonal antibody to c-RAF (Phospho-S621) Immunogen: KLH-conjugated synthetic phosphopeptide corresponding to residues surrounding S621 of human c-RAF protein. The exact sequence is proprietary. Purification: The antibody was purified by immunogen affinity chromatography. Clonality: Polyclonal Form: Liquid in 0.42% Potassium phosphate, 0.87% Sodium chloride, pH 7.3, 30% glycerol, and 0.01% sodium azide. Dilution: WB (1/500 - 1/1000), IH (1/50 - 1/200), IF/IC (1/100 - 1/500) Gene Symbol: RAF1 Alternative Names: RAF; RAF proto-oncogene serine/threonine-protein kinase; Proto-oncogene c-RAF; cRaf; Raf-1Entrez Gene (Human): 5894Entrez Gene (Mouse) : 110157Entrez Gene (Rat) : 24703SwissProt (Human): P04049SwissProt (Mouse) : Q99N57SwissProt (Rat) : P11345Storage/Stability : Shipped at 4°C. Upon delivery aliquot and store at -20°C for one year. Avoid freeze/thaw cycles.
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Western blot analysis of c-RAF (Phospho-S621) expression in Panc1 (A), HEK293T (B), PC3 (C) whole cell lysates.
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Immunohistochemical analysis of c-RAF (Phospho-S621) staining in human heart formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
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Immunofluorescent analysis of c-RAF (Phospho-S621) staining in HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 °C in a hidified chamber. Cells were washed with PBST and incubated with a Alexa Fluor 594-conjugated secondary antibody (red) in PBS at room temperature in the dark.