Product Name :
Raf-1 (Phospho-Ser621) polyclonal antibody 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. Activation of c-Raf is the best understood and involves phosphorylation at multiple activating sites, including Ser338, Tyr341, Thr491, Ser494, Ser497, and Ser499. p21-activated kinase (PAK) has been shown to phosphorylate c-Raf at Ser338, and the Src family phosphorylates Tyr341 to induce c-Raf activity. 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. Inhibitory 14-3-3 binding sites on c-Raf (Ser259 and Ser621) can be phosphorylated by Akt and AMPK, respectively. While A-Raf, B-Raf, and c-Raf are similar in sequence and function, differential regulation has been observed. 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. Research studies have shown that the B-Raf mutation V600E results in elevated kinase activity and is commonly found in malignant melanoma. 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. Product :
Rabbit IgG, 1mg/ml in PBS with 0.02% sodium azide, 50% glycerol, pH7.2. Storage&Stability :
Store at 4°C short term. Aliquot and store at -20°C long term. Avoid freeze-thaw cycles. Specificity :
Raf-1 (Phospho-Ser621) polyclonal antibody detects endogenous levels of Raf-1 protein only when phosphorylated at Ser621. Immunogen :
Synthetic phosphopeptide derived from human Raf-1 around the phosphorylation site of Serine 621. Conjugate :
Unconjugated Modification :
Phosphorylated

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  Western blot (WB) analysis of Raf-1 (Phospho-Ser621) polyclonal antibody at 1:1000 dilution Lane1:Hela whole cell lysate(30ug) Lane2:Hela treated with λ-phosphatase whole cell lysate(30ug) Lane3:CT-26 whole cell lysate(30ug) Lane4:PC12 whole cell lysate(30ug)
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  Immunofluorescence analysis of EC9706 cells using Raf-1 (Phospho-Ser621) pAb at dilution of 1:200 (40x lens).

Bioworld Biotech only provide peptides for our antibodies and do not provide additional peptide customization services.

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Price/Size :

USD 368/1mg/vial

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Tips: 

For phospho antibody, we provide phospho peptide（0.5mg) and non-phospho peptide(0.5mg).

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Describe :

Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. This mechanism is useful when non-specific binding is an issue, for example, in Western blotting (WB) and Immunohistochemistry (IHC). By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific; Specific binding will be absent from the western blot or IHC performed with the neutralized antibody.

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Formula:

Synthetic peptide was lyophilized with 100% acetonitrile and is supplied as a powder. Reconstitute with 0.1 ml DI water for a final concentration of 10 mg/ml.The purity is >90%,tested by HPLC and MS.

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Storage:

The freeze-dried powder is more stable. For short time at 2-8°C. For long term storage store at -20°C. 

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Note :

This product is for research use only (RUO only). Not for use in diagnostic or therapeutic procedures.

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