|Application ||WB, IHC|
|Reactivity||Human, Mouse, Rat|
|Calculated MW||73052 Da|
|Application & Usage||Western blotting (0.5-4 µg/ml) and Immunohistochemistry (20-40 µg/ml). However, the optimal conditions should be determined individually. The antibody detects ~74 kDa c-Raf when phosphorylated at Ser259.|
|Other Names||RAF1 , cRaf , Raf-1 , c-Raf , RAF , CRAF , C-RAF, RAF proto-oncogene serine/threonine-protein kinase; Proto-oncogene c-RAF; cRaf; Raf-1|
|Formulation||100 µg (0.5 mg/ml) affinity purified rabbit anti-Phospho-Raf (Ser259) polyclonal antibody in phosphate buffered saline (PBS), pH 7.2, containing 50% glycerol, 1% BSA, 0.02% thimerosal.|
|Handling||The antibody solution should be gently mixed before use.|
|Reconstitution & Storage||-20 °C|
|Precautions||Phospho-Raf Antibody is for research use only and not for use in diagnostic or therapeutic procedures.|
|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.|
|Cellular Location||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 Location||In skeletal muscle, isoform 1 is more abundant than isoform 2.|
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Provided below are standard protocols that you may find useful for product applications.
c-Raf is a mitogen-activated protein kinase. Binding of Raf to 14-3-3 is required for c-Raf kinase activity, and also blocks the ability of phosphatase to inhibit c-Raf. 14-3-3 binds c-Raf at two phospho-serine residues: Ser259 and Ser621. Mutation of Ser259 to alanine constitutively activates Raf kinase activity, but mutation of Ser621 to alanine results in an inactive kinase.
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