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Anti-Phospho-Raf-1 (S259) antibody

[STJ90399] Download PDF Print Data Sheet

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Product name Anti-Phospho-Raf-1 (S259) antibody
Short Description Rabbit Polyclonal to Phospho-Raf-1 (S259).
Description Raf-1 is a protein encoded by the RAF1 gene which is approximately 73 kDa. Raf-1 is localised to the cytoplasm, cell membrane, mitochondrion and nucleus. It is involved in RET signalling, the IL-2 pathway, regulation of lipid metabolism and insulin signalling-generic cascades. When it is activated, the cellular RAF1 protein can phosphorylate to activate the dual specificity protein kinases MEK1 and MEK2, which in turn phosphorylates to activate the serine/threonine specific protein kinases, ERK1 and ERK2. It plays an important role in the control of gene expression involved in the cell division cycle, apoptosis, cell differentiation and cell migration. Raf-1 is expressed in skeletal muscle with isoform 1 is more abundant than isoform 2. Mutations in the RAF1 gene result in Noonan syndrome, Leopard syndrome and cardiomyopathy. The antibody STJ90399 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen. This primary antibody specifically binds to endogenous Raf-1 protein which only binds about S259 when S259 is phosphorylated.
Applications IHC-p, ELISA, WB
Dilution range WB 1:500-1:2000 / IHC 1:100-1:300 / ELISA 1:20000 /
Specificity Phospho-Raf-1 (S259) Polyclonal Antibody detects endogenous levels of Raf-1 protein only when phosphorylated at S259.
Protein Name RAF proto-oncogene serine/threonine-protein kinase - RAF1 - Raf-1 - c-Raf - CRAF - Raf-1 proto-oncogene - serine/threonine kinase
Immunogen Synthesized peptide derived from human Raf-1 around the phosphorylation site of S259.
Immunogen Region 200-280 aa
Storage Instruction Store at -20°C, and avoid repeat freeze-thaw cycles.
Note For Research Use Only (RUO).
Host Rabbit
Clonality Polyclonal
Reactivity Human, Monkey, Mouse, Rat
Conjugation Unconjugated
Concentration 1 mg/ml
Purification The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.
Isotype IgG
Formulation Liquid in PBS containing 50% glycerol, 0.5% BSA and 0.02% sodium azide.
Gene ID 5895 (Human) / 110157 (Mouse) / 24703 (Rat)
Gene Symbol RAF1
Molecular Weight 73 kDa
Database Links Human UniProt/Swiss-Prot:P04049 Mouse UniPort/Swiss-Prot: Q99N57 Rat UniProt/Swiss-Port: P11345 Human Entrez Gene: 5895 Mouse Entrez Gene: 110157 Rat Entrez Gene: Rn.33262
Alternative Names RAF1 / Raf-1 proto-oncogene / serine /threonine kinase / A306_15425 antibody / Anapl_09074 antibody / AS27_00394 antibody / AS28_07522 antibody / BRAF antibody / CB1_000465013 antibody / CMD1NN antibody / c-mil protein antibody / c-Raf antibody / CRAF antibody / Craf1 antibody / Craf1 transforming antibody / C-Raf proto-oncogene / serine /threonine kinase antibody / D623_10028290 antibody / GB16508 antibody / GW7_06109 antibody / H920_11160 antibody / hypothetical protein antibody / M91_19148 antibody / M959_11320 antibody / MDA_GLEAN10023754 antibody / MIL proto-oncogene serine /threonine-protein kinase antibody / murine leukemia viral (v-raf-1) oncogene homolog 1 antibody / murine leukemia viral (v-raf-1) oncogene homolog 1 (3611-MSV) antibody / murine sarcoma 3611 oncogene 1 antibody / N300_10964 antibody / N301_14215 antibody / N302_07201 antibody / N303_00462 antibody / N305_01579 antibody / N306_08254 antibody / N307_05496 antibody / N308_01932 antibody / N309_15005 antibody / N310_13119 antibody / N311_12207 antibody / N320_09181 antibody / N321_12663 antibody / N324_04212 antibody / N327_08178 antibody / N329_10964 antibody / N330_06064 antibody / N331_01439 antibody / N332_13045 antibody / N333_09715 antibody / N334_11976 antibody / N335_10335 antibody / N336_09007 antibody / N340_03049 antibody / N341_03296 antibody / NS5 antibody / Oncogene RAF1 antibody / PAL_GLEAN10022307 antibody / PANDA_010199 antibody / proto-oncogene c-RAF antibody / RAF-1 antibody / raf1c antibody / Raf kinase / effector of Ras antibody / raf proto-oncogene serine /threonine protein kinase antibody / RAF proto-oncogene serine /threonine-protein kinase antibody / RAF proto-oncogene serine /threonine-protein kinase-like protein antibody / sarcoma 3611 oncogene antibody / serine /threonine protein kinase antibody / serine /threonine protein kinase RAF1 antibody / serine /threonine protein kinase RAF1c antibody / TREES_T100019833 antibody / UY3_03909 antibody / v-Raf antibody / v-raf-1 leukemia viral oncogene 1 antibody / v-raf-1 murine leukemia viral oncogene homolog 1 antibody / v-raf-1 murine leukemia viral oncogene-like protein 1 antibody / v-raf-1 murine leukemia viral oncoprotein-like protein 1 antibody / v-raf-leukemia viral oncogene 1 antibody / v-raf murine sarcoma viral oncogene homolog B antibody / v-raf murine sarcoma viral oncogene-like protein B antibody / Y1Q_012506 antibody / Y956_05358 antibody / Z169_12605 antibody
Function Serine/threonine-protein kinase that acts as a regulatory link between the membrane-associated Ras GTPases and the MAPK/ERK cascade, and this critical regulatory link functions as a switch determining cell fate decisions including proliferation, differentiation, apoptosis, survival and oncogenic transformation. RAF1 activation initiates a mitogen-activated protein kinase (MAPK) cascade that comprises a sequential phosphorylation of the dual-specific MAPK kinases (MAP2K1/MEK1 and MAP2K2/MEK2) and the extracellular signal-regulated kinases (MAPK3/ERK1 and MAPK1/ERK2). The phosphorylated form of RAF1 (on residues Ser-338 and Ser-339, by PAK1) phosphorylates BAD/Bcl2-antagonist of cell death at 'Ser-75'. Phosphorylates adenylyl cyclases: ADCY2, ADCY5 and ADCY6, resulting in their activation. Phosphorylates PPP1R12A resulting in inhibition of the phosphatase activity. Phosphorylates TNNT2/cardiac muscle troponin T. Can promote NF-kB activation and inhibit signal transducers involved in motility (ROCK2), apoptosis (MAP3K5/ASK1 and STK3/MST2), proliferation and angiogenesis (RB1). Can protect cells from apoptosis also by translocating to the mitochondria where it binds BCL2 and displaces BAD/Bcl2-antagonist of cell death. Regulates Rho signaling and migration, and is required for normal wound healing. Plays a role in the oncogenic transformation of epithelial cells via repression of the TJ protein, occludin (OCLN) by inducing the up-regulation of a transcriptional repressor SNAI2/SLUG, which induces down-regulation of OCLN. Restricts caspase activation in response to selected stimuli, notably Fas stimulation, pathogen-mediated macrophage apoptosis, and erythroid differentiation.
Post-translational Modifications 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.
Cellular Localization Cytoplasm. Cell membrane. Mitochondrion. Nucleus. Note=Colocalizes with RGS14 and BRAF in both the cytoplasm and membranes. Phosphorylation at Ser-259 impairs its membrane accumulation. Recruited to the cell membrane by the active Ras protein. Phosphorylation at Ser-338 and Ser-339 by PAK1 is required for its mitochondrial localization. Retinoic acid-induced Ser-621 phosphorylated form of RAF1 is predominantly localized at the nucleus.
Tissue Specificity In skeletal muscle, isoform 1 is more abundant than isoform 2.


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