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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-A-Raf (Ser299) Antibody 4431 40 µl
H M R 68 Rabbit 
A-Raf Antibody 4432 40 µl
H M R 68 Rabbit 
Phospho-c-Raf (Ser338) (56A6) Rabbit mAb 9427 40 µl
H M R Mk 74 Rabbit IgG
Phospho-c-Raf (Ser289/296/301) Antibody 9431 40 µl
H M 74 Rabbit 
Phospho-c-Raf (Ser259) Antibody 9421 40 µl
H M R X 74 Rabbit 
c-Raf (D5X6R) Mouse mAb 12552 40 µl
H M R Mk B Pg 75 Mouse IgG1
Phospho-B-Raf (Ser445) Antibody 2696 40 µl
H M R Mk 86 Rabbit 
B-Raf (55C6) Rabbit mAb 9433 40 µl
H M R Mk 86 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
All Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl
All Horse 

Product Description

The Raf Family Antibody Sampler Kit provides a fast and economical means to investigate Raf signaling. The kit contains enough primary and secondary antibody to perform four Western blot experiments.

Specificity / Sensitivity

Each antibody in the Raf Family Antibody Sampler Kit recognizes only its specific target and does not cross react with other Raf family members.

Source / Purification

The phospho-specific polyclonal antibodies are produced by immunizing rabbits with a synthetic phosphopeptide corresponding to residues surrounding Ser299 of human A-Raf, Ser445 of human B-Raf and Ser259, 289, 296 and 301 of c-Raf. The total polyclonal antibody is produced by immunizing rabbits with a synthetic peptide corresponding to residues close to the linker domain of human A-Raf. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. The monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser338 of human c-Raf, a synthetic peptide corresponding to human B-Raf and a recombinant protein corresponding to residues in the middle of human c-Raf protein.

A-Raf, B-Raf, and c-Raf (Raf-1) are the main effectors recruited by GTP-bound Ras to activate the MEK-MAP kinase pathway (1). Activation of c-Raf is the best understood and involves phosphorylation at multiple activating sites including Ser338, Tyr341, Thr491, Ser494, Ser497, and Ser499 (2). p21-activated protein kinase (PAK) has been shown to phosphorylate c-Raf at Ser338 and the Src family phosphorylates Tyr341 to induce c-Raf activity (3,4). Ser338 of c-Raf corresponds to similar sites in A-Raf (Ser299) and B-Raf (Ser445), although this site is constitutively phosphorylated in B-Raf (5). Inhibitory 14-3-3 binding sites on c-Raf (Ser259 and Ser621) can be phosphorylated by Akt and AMPK, respectively (6,7). While A-Raf, B-Raf, and c-Raf are similar in sequence and function, differential regulation has been observed (8). Of particular interest, B-Raf contains three consensus Akt phosphorylation sites (Ser364, Ser428, and Thr439) and lacks a site equivalent to Tyr341 of c-Raf (8,9). Research studies have shown that the B-Raf mutation V600E results in elevated kinase activity and is commonly found in malignant melanoma (10). Six residues of c-Raf (Ser29, Ser43, Ser289, Ser296, Ser301, and Ser642) become hyperphosphorylated in a manner consistent with c-Raf inactivation. The hyperphosphorylation of these six sites is dependent on downstream MEK signaling and renders c-Raf unresponsive to subsequent activation events (11).

1.  Avruch, J. et al. (1994) Trends Biochem Sci 19, 279-83.

2.  Chong, H. et al. (2001) EMBO J 20, 3716-27.

3.  King, A.J. et al. (1998) Nature 396, 180-3.

4.  Fabian, J.R. et al. (1993) Mol Cell Biol 13, 7170-9.

5.  Mason, C.S. et al. (1999) EMBO J 18, 2137-48.

6.  Sprenkle, A.B. et al. (1997) FEBS Lett 403, 254-8.

7.  Marais, R. et al. (1997) J Biol Chem 272, 4378-83.

8.  Zimmermann, S. and Moelling, K. (1999) Science 286, 1741-4.

9.  Guan, K.L. et al. (2000) J Biol Chem 275, 27354-9.

10.  Davies, H. et al. (2002) Nature 417, 949-54.

11.  Dougherty, M.K. et al. (2005) Mol Cell 17, 215-24.

Entrez-Gene Id 369, 673, 5894
Swiss-Prot Acc. P10398, P15056, P04049

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 5,675,063.