General description
Note: 1 T = 1 test. Sufficient for 10 Western miniblots.
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 Tyr340/341.
Recognizes c-Raf phosphorylated at Tyr340 and Tyr341 in HEK293 cells overexpressing c-Raf.
This PhosphoDetect
Immunogen
Human
a synthetic phosphopeptide corresponding to amino acids surrounding the Tyr³⁴⁰ and Tyr³⁴¹, phosphorylation sites of human c-Raf (Raf-1)
Application
Immunoblotting (1:1000)
Immunocytochemistry (see comments)
Physical form
In phosphate buffer (without Mg2+ and Ca2+), 1 mg/ml BSA, 50% glycerol, pH 7.3 (+/- 0.1).
Preparation Note
Following initial thaw, aliquot and freeze (-20°C).
Analysis Note
Positive Control
HEK293 cells over expressing c-Raf, phosphorylated in vitro by Src.
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.
Legal Information
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Toxicity: Standard Handling (A)
| biological source | rabbit |
| Quality Segment | 100 |
| antibody form | affinity isolated antibody |
| antibody product type | primary antibodies |
| clone | polyclonal |
| form | liquid |
| contains | ≤0.1% sodium azide as preservative |
| species reactivity | rat, human, mouse |
| manufacturer/tradename | Calbiochem® |
| storage condition | OK to freeze,avoid repeated freeze/thaw cycles |
| isotype | IgG |
| shipped in | wet ice |
| storage temp. | −20°C |
| target post-translational modification | phosphorylation (pTyr340/pTyr341) |
| Gene Information | human ... RAF1(5894) |
