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c-Raf (D5X6R) Mouse mAb #12552

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  • WB
Western Blotting Image 1: c-Raf (D5X6R) Mouse mAb
Western blot analysis of extracts from various cell lines using c-Raf (D5X6R) Mouse mAb.

To Purchase # 12552

Supporting Data

REACTIVITY H M R Mk B Pg
SENSITIVITY Endogenous
MW (kDa) 75
Source/Isotype Mouse IgG1
Application Key:
  • WB-Western Blotting 
Species Cross-Reactivity Key:
  • H-Human 
  • M-Mouse 
  • R-Rat 
  • Mk-Monkey 
  • B-Bovine 
  • Pg-Pig 
  • Related Products

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Protocol

Specificity / Sensitivity

c-Raf (D5X6R) Mouse mAb recognizes endogenous levels of total c-Raf protein. This antibody does not cross-react with A-Raf or B-Raf.

Species Reactivity:

Human, Mouse, Rat, Monkey, Bovine, Pig

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein corresponding to residues in the middle of human c-Raf protein.

Background

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 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).

Pathways

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For Research Use Only. Not for Use in Diagnostic Procedures.
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