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Raf-1 (Phospho-Ser621) polyclonal antibody

Product Specifications

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.

CAS Number

9007-83-4

Structure Composition

Rabbit IgG, 1mg/ml in PBS with 0.02% sodium azide, 50% glycerol, pH7.2.

Product Name Alternative

RAF proto-oncogene serine/threonine-protein kinase; Proto-oncogene c-RAF; cRaf; Raf-1; RAF1; RAF

Swiss Prot

P04049

Reactivity

Human, Rat, Mouse

Immunogen

Synthetic phosphopeptide derived from human Raf-1 around the phosphorylation site of Serine 621.

Conjugation

Unconjugated

Related Pathways

ESC Pluripotency and Differentiation Signaling Pathway, ErbB HER Signaling, G Protein-coupled Receptors Signaling to MAPK Erk, MAPK Erk in Growth and Differentiation Pathway (, Jak Stat : IL-6 Receptor Signaling, Inhibition of Apoptosis, Insulin &Glucose Signaling, T cell receptor signaling

Applications

WB IF IP

Dilution

WB: 1:1000~1:2000 IF: 1:50~1:200 IP: 1:50~1:200

Purity

The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen and the purity is > 95% (by SDS-PAGE) .

Modification

Phosphorylated

Molecular Weight

~ 75 kDa

Storage Conditions

Store at 4°C short term. Aliquot and store at -20°C long term. Avoid freeze-thaw cycles.

Notes

For research use only, not for use in diagnostic procedure.

Specificity

Raf-1 (Phospho-Ser621) polyclonal antibody detects endogenous levels of Raf-1 protein only when phosphorylated at Ser621.

Applications Notes

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)

Host or Source

Rabbit

Available Sizes

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