Raf Family, Antibody Sampler Kit, BioAssay(TM)

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). 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).\n\nKit Includes:\nQuantity\nApplications\nReactivity\nMW (kD)\n\n(1) Rabbit x Phospho-A-Raf (Ser299) Antibody\n40ul\nWB\nHuman Mouse (Rat)\n68kD\nRabbit\n\n(2) Rabbit x A-Raf Antibody\n40ul\nWB IP\nHuman Mouse Rat\n68kD\n\n(3) Rabbit x Phospho-c-Raf (Ser338) (mAb)\n40ul\nWB\nHuman Mouse Rat Monkey\n74kD\n\n(4) Rabbit x Phospho-c-Raf (Ser289/296/301)\n40ul\nWB\nHuman Mouse\n74kD\n\n(5) Rabbit x Phospho-c-Raf (Ser259)\n40ul\nWB IP\nHuman Mouse Rat Xenopus (Chicken)\n74kD\n\n(6) Rabbit x c-Raf Antibody\n40ul\nWB IP\nHuman Mouse Rat\n65 to 75kD\n\n(7) Rabbit x Phospho-B-Raf (Ser445)\n40ul\nWB\nHuman Mouse Rat Monkey (Chicken)\n86kD\n\n(8) Mouse x B-Raf (mAb)\n40ul\nWB IP FC\nHuman Mouse Rat Monkey\n86kD

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SPECIFICATIONS

Catalog Number

R0495-12

Size

1Kit

Applications

WB

Purity

Polyclonal antibodies are purified by Protein A and immunoaffinity chromatography.

References

1 Avruch, J. et al. (1994) Trends Biochem. Sci. 19, 279-283. \n 2 Chong, H. et al. (2001) EMBO J. 20, 3716-3727. \n 3 King, A.J. et al. (1998) Nature 396, 180-183. \n 4 Fabian, J.R. et al. (1993) Mol. Cell Biol. 13, 7170-7179. \n 5 Mason, C.S. et al. (1999) EMBO J. 18, 2137-2148. \n 6 Zimmermann, S. and Moelling, K. (1999) Science 286, 1741-1744. \n 7 Sprenkle, A.B. et al. (1997) FEBS Lett. 403, 254-258. \n 8 Marais, R. et al. (1997) J. Biol. Chem. 272, 4378-4383. \n 9 Guan, K.L. et al. (2000) J. Biol. Chem. 275, 27354-27359. \n 10 Davies, H. et al. (2002) Nature 417, 949-954. \n 11 Dougherty, M.K. et al. (2005) Mol. Cell 17, 215-224.

Additional Info

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