Analysis Note

Positive ControlHEK293 cells over expressing c-Raf, phosphorylated in vitro by Src.

Application

Immunoblotting (1:1000)r>Immunocytochemistry (see comments)

General description

This PhosphoDetect™ Anti-c-Raf (pTyr³⁴⁰, Tyr³⁴¹) Rabbit pAb is validated for use in WB, ICC for the detection of c-Raf (pTyr³⁴⁰, Tyr³⁴¹).

Rabbit polyclonal antibody adsorbed against non-phosphopeptide corresponding to the immunogen phosphorylation site, followed by immunoaffinity chromatography. Recognizes the ~74 KDa c-Raf protein phosphorylated on Tyr^340/341.

Note: 1 T = 1 test. Sufficient for 10 Western miniblots.

Recognizes c-Raf phosphorylated at Tyr³⁴⁰ and Tyr³⁴¹ in HEK293 cells overexpressing c-Raf.

Immunogen

a synthetic phosphopeptide corresponding to amino acids surrounding the Tyr³⁴⁰ and Tyr³⁴¹, phosphorylation sites of human c-Raf (Raf-1)

Human

Legal Information

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

The immunogen sequence is 100% conserved in mouse and rat, but cross-reactivity has not been tested. This antibody has also been reported to work for immunocytochemistry. Variables associated with assay conditions will dictate the proper working dilution.

Yan, F., and Polk, D.B. 2001. Cancer Res.61, 963.Yip-Schneider, M.T., et al. 2000. Biochem. J.351, 151.Mason, C.S. et al. 1999. EMBO J.18, 2137.Bonner, T.I. et al. 1985. Mol. Cell. Biol.5, 1400.Moelling, K. et al. 1984. Nature312, 558.Rapp, U.R. et al. 1983. Proc. Natl. Acad. Sci. USA80, 4218.

Packaging

10 tests in Plastic ampoule

Physical form

In phosphate buffer (without Mg²⁺ and Ca²⁺), 1 mg/ml BSA, 50% glycerol, pH 7.3 (+/- 0.1).

Reconstitution

Following initial thaw, aliquot and freeze (-20°C).

Warning

Toxicity: Standard Handling (A)