ARG52409
anti-Raf1 phospho (Ser301) antibody
anti-Raf1 phospho (Ser301) antibody for Western blot and Rat
Cancer antibody; Cell Biology and Cellular Response antibody; Cell Death antibody; Metabolism antibody; Signaling Transduction antibody
Overview
| Product Description | Rabbit Polyclonal antibody recognizes Raf1 phospho (Ser301) |
|---|---|
| Tested Reactivity | Rat |
| Predict Reactivity | Ms, Bov, Chk, Dog, NHuPrm, Xenopus laevis |
| Tested Application | WB |
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Target Name | Raf1 |
| Antigen Species | Rat |
| Immunogen | Synthetic phospho-peptide corresponding to amino acid residues surrounding Ser301 conjugated to KLH |
| Conjugation | Un-conjugated |
| Alternate Names | c-Raf; cRaf; Proto-oncogene c-RAF; CRAF; RAF proto-oncogene serine/threonine-protein kinase; CMD1NN; Raf-1; EC 2.7.11.1; NS5 |
Application Instructions
| Application Suggestion |
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| Application Note | Specific for the ~74k Raf-1 protein phosphorylated at Ser301. Immunolabeling of the Raf-1 protein band is blocked by λ-phosphatase treatment. * The dilutions indicate recommended starting dilutions and the optimal dilutions or concentrations should be determined by the scientist. |
Properties
| Form | Liquid |
|---|---|
| Purification | Affinity Purified |
| Buffer | 10 mM HEPES (pH 7.5), 150 mM NaCl, 0.1 mg/ml BSA and 50% Glycerol |
| Stabilizer | 0.1 mg/ml BSA, 50% Glycerol |
| Storage Instruction | For continuous use, store undiluted antibody at 2-8°C for up to a week. For long-term storage, aliquot and store at -20°C. Storage in frost free freezers is not recommended. Avoid repeated freeze/thaw cycles. Suggest spin the vial prior to opening. The antibody solution should be gently mixed before use. |
| Note | For laboratory research only, not for drug, diagnostic or other use. |
Bioinformation
| Database Links |
Swiss-port # P11345 Rat RAF proto-oncogene serine/threonine-protein kinase |
|---|---|
| Gene Symbol | RAF1 |
| Gene Full Name | v-raf-leukemia viral oncogene 1 |
| Background | The Ras pathway is a critical signal transduction cascade involved in regulating cellular proliferation, differentiation, survival, and oncogenic transformation. Members of the Raf serine/threonine kinase family are key intermediates in this cascade, functioning to relay signals from activated Ras to the downstream protein kinases MEK and ERK (Marshall, 1996). Previous studies have shown that phosphorylation is required for Raf-1 activation (Dhillon and Kolch, 2002; Chong et al., 2003). Recent work has demonstrated that phosphorylation also regulates the downregulation of Raf (Dougherty et al., 2005) with two sites participating: Ser301 and Ser642. |
| Research Area | Cancer antibody; Cell Biology and Cellular Response antibody; Cell Death antibody; Metabolism antibody; Signaling Transduction antibody |
| Calculated MW | 73 kDa |
| PTM | Phosphorylation at Thr-269, Ser-338, Tyr-341, Thr-491 and Ser-494 results in its activation. Phosphorylation at Ser-29, Ser-43, Ser-289, Ser-296, Ser-301 and Ser-642 by MAPK1/ERK2 results in its inactivation. Phosphorylation at Ser-259 induces the interaction with YWHAZ and inactivates kinase activity. Dephosphorylation of Ser-259 by the complex containing protein phosphatase 1, SHOC2 and M-Ras/MRAS relieves inactivation, leading to stimulate RAF1 activity. Phosphorylation at Ser-338 by PAK1 and PAK5 and Ser-339 by PAK1 is required for its mitochondrial localization. Phosphorylation at Ser-621 in response to growth factor treatment stabilizes the protein, possibly by preventing proteasomal degradation. Phosphorylation at Ser-289, Ser-296, Ser-301, Ser-338 and Ser-621 are somehow linked to the methylation potential of cells. Treatment of cells with HGF in the presence of the methylation inhibitor 5'-methylthioadenosine (MTA) results in increased phosphorylation at Ser-338 and Ser-621 and decreased phosphorylation at Ser-296, Ser-301 and Ser-338. Dephosphorylation at Ser-338 by PPP5C results in a activity decrease. Methylated at Arg-563 in response to EGF treatment. This modification leads to destabilization of the protein, possibly through proteasomal degradation. |
Images (1) Click the Picture to Zoom In
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ARG52409 anti-Raf1 phospho (Ser301) antibody WB image
Western blot: UV treated Human Jurkat cell lysate stained with ARG52409 anti-Raf1 phospho (Ser301) antibody showing specific immunolabeling of the ~74k Raf-1 protein (Control). The phosphospecificity of this labeling is shown in the second lane (lambda-phosphatase: λ-Ptase). The blot is identical to the control except that it was incubated in λ-Ptase (1200 units for 30 min) before being exposed to the phospho Ser301 Raf-1 antibody. The immunolabeling of Raf-1 is completely eliminated by treatment with λ-Ptase.
