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c-Raf (D5X6R) Mouse Monoclonal Antibody (BSA and Azide Free) #85554

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  • WB
  • ELISA+

    Product Specifications

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

    Product Information

    Product Usage Information

    This product is the carrier free version of product #12552. All data were generated using the same antibody clone in the standard formulation which contains BSA and glycerol.

    This formulation is ideal for use with technologies requiring specialized or custom antibody labeling, including fluorophores, metals, lanthanides, and oligonucleotides. It is not recommended for ChIP, ChIP-seq, CUT&RUN or CUT&Tag assays. If you require a carrier free formulation for chromatin profiling, please contact us. Optimal dilutions/concentrations should be determined by the end user.

    BSA and Azide Free antibodies are quality control tested by size exclusion chromatography (SEC) to determine antibody integrity.

    Formulation

    Supplied in 1X PBS (10 mM Na2HPO4, 3 mM KCl, 2 mM KH2PO4, and 140 mM NaCl (pH 7.8)). BSA and Azide Free.

    For standard formulation of this product see product #12552

    Storage

    Store at -20°C. This product will freeze at -20°C so it is recommended to aliquot into single-use vials to avoid multiple freeze/thaw cycles. A slight precipitate may be present and can be dissolved by gently vortexing. This will not interfere with antibody performance.

    Specificity / Sensitivity

    c-Raf (D5X6R) Mouse Monoclonal Antibody (BSA and Azide Free) 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 (Ser446), 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 (Ser365, Ser429, and Thr440) 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).

    Alternate Names

    c-Raf; C-Raf proto-oncogene, serine/threonine kinase; CMD1NN; cRaf; NS5; Oncogene RAF1; Proto-oncogene c-RAF; RAF; raf proto-oncogene serine/threonine protein kinase; RAF proto-oncogene serine/threonine-protein kinase; Raf-1; Raf-1 proto-oncogene, serine/threonine kinase; RAF1; v-raf-1 murine leukemia viral oncogene homolog 1; v-raf-1 murine leukemia viral oncogene-like protein 1

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