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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
Proto-oncogene c-RAF
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 5894
Gene Symbol RAF1
Molecular Weight 73 kDa
Database Links HGNC:9829
Alternative Names RAF1 antibody
RAF antibody
RAF proto-oncogene serine/threonine-protein kinase antibody
Proto-oncogene c-RAF antibody
cRaf antibody
Raf-1 antibody
Phospho-Raf-1 (S259) 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.
Swiss-Prot Key P04049_HUMAN


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