|Application ||WB, IHC|
|Reactivity||Human, Mouse, Rat|
|Calculated MW||73052 Da|
|Other Names||RAF proto-oncogene serine/threonine-protein kinase, Proto-oncogene c-RAF, cRaf, Raf-1, RAF1, RAF|
|Target/Specificity||A synthetic peptide corresponding to residues surrounding Serine 338-339 of human Raf was used as immunogen.|
|Format||50 mM Tris-Glycine (pH 7.4), 0.15 M NaCl, 40% Glycerol, 0.01% sodium azide and 0.05% BSA.|
|Storage||Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.|
|Precautions||Raf-1 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.
Raf-1 (c-Raf) is a protein kinase that is involved in the transduction of mitogenic signals from the cell membrane to the nucleus. Raf-1 is recruited by and binds to GTP-bound Ras, activating the MEK-MAP kinase pathway (1). Raf-1 is activated via rapid hyperphosphorylation at multiple sites in mammalian cells, including Ser43, Ser259, and Ser621 (2), as well as Ser338, Tyr341, Thr491, and Ser494 (3). p21-activated protein kinase (PAK) phosphorylates Raf-1 at Ser338 in response to Ras, Src and epidermal growth factor (4).
1. Avruch, J., et al. Raf meets Ras: completing the framework of a signal transduction pathway. Trends Biochem. Sci. 19: 279
2. Morrison, D.K., et al. Identification of the major phosphorylation sites of the Raf-1 kinase. J Biol Chem. 268: 17309
3. Chong, H., et al. Positive and negative regulation of Raf kinase activity and function by phosphorylation. EMBO J. 20: 3716
4. King, A.J., et al. The protein kinase Pak3 positively regulates Raf-1 activity through phosphorylation of serine 338. Nature 396: 180
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