Western Blot (WB)
(Western blot analysis of extracts of HeLa and 293T cells, using Phospho-RAF1-S621 antibody at 1:1000 dilution. HeLa cells were treated by EGF (100ng/ml) for 30 minutes after serum-starvation overnight. 293T cells were treated by PMA/TPA (200nM) for 30 minutes or treated by EGF (25ug/ml) for 30 minutes after serum-starvation overnight.Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (MBS128200) at 1:10000 dilution.Lysates/proteins: 25ug per lane.Blocking buffer: 3% BSA.)
Rabbit anti-Human RAF1-S621 Antibody | anti-RAF1 antibody
Phospho-RAF1-S621 Polyclonal Antibody
IF: 1:50 - 1:200
Buffer: PBS with 0.02% sodium azide, 50% glycerol, pH7.3.
Western Blot (WB)
(Western blot analysis of extracts of HeLa and 293T cells, using Phospho-RAF1-S621 antibody at 1:1000 dilution. HeLa cells were treated by EGF (100ng/ml) for 30 minutes after serum-starvation overnight. 293T cells were treated by PMA/TPA (200nM) for 30 minutes or treated by EGF (25ug/ml) for 30 minutes after serum-starvation overnight.Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (MBS128200) at 1:10000 dilution.Lysates/proteins: 25ug per lane.Blocking buffer: 3% BSA.)
Western Blot (WB)
(Western blot analysis of extracts of HeLa and 293T cells, using Phospho-RAF1-S621 antibody at 1:1000 dilution. HeLa cells were treated by EGF (100ng/ml) for 30 minutes after serum-starvation overnight. 293T cells were treated by PMA/TPA (200nM) for 30 minutes or treated by EGF (25ug/ml) for 30 minutes after serum-starvation overnight.Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (MBS128200) at 1:10000 dilution.Lysates/proteins: 25ug per lane.Blocking buffer: 3% BSA.)
Immunofluorescence (IF)
(Immunofluorescence analysis of U2OS cells using Phospho-RAF1-S621 antibody. Blue: DAPI for nuclear staining.)
Immunofluorescence (IF)
(Immunofluorescence analysis of U2OS cells using Phospho-RAF1-S621 antibody. Blue: DAPI for nuclear staining.)
Immunoprecipitation (IP)
(Immunoprecipitation analysis of 200ug extracts of HeLa cells treated by EGF using 2.5ug Phospho-RAF1-S621 antibody. Western blot was performed from the immunoprecipitate using Phospho-RAF1-S621 antibody at a dilition of 1:1000.)
Immunoprecipitation (IP)
(Immunoprecipitation analysis of 200ug extracts of HeLa cells treated by EGF using 2.5ug Phospho-RAF1-S621 antibody. Western blot was performed from the immunoprecipitate using Phospho-RAF1-S621 antibody at a dilition of 1:1000.)
NCBI and Uniprot Product Information
NCBI Description
This gene is the cellular homolog of viral raf gene (v-raf). The encoded protein is a MAP kinase kinase kinase (MAP3K), which functions downstream of the Ras family of membrane associated GTPases to which it binds directly. Once activated, the cellular RAF1 protein can phosphorylate to activate the dual specificity protein kinases MEK1 and MEK2, which in turn phosphorylate to activate the serine/threonine specific protein kinases, ERK1 and ERK2. Activated ERKs are pleiotropic effectors of cell physiology and play an important role in the control of gene expression involved in the cell division cycle, apoptosis, cell differentiation and cell migration. Mutations in this gene are associated with Noonan syndrome 5 and LEOPARD syndrome 2. [provided by RefSeq, Jul 2008]
Uniprot Description
RAF1: a proto-oncogenic TKL kinase of the RAF family. Functions downstream of the Ras family of membrane associated GTPases to which it binds directly. Once activated Raf-1 can phosphorylate and activate MEK1/2, which in turn phosphorylate and activate ERK1/2. 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 (MEK1/2) and the extracellular signal-regulated kinases (ERK1/2). RAF1, subsequent to phosphorylation by PAK1, phosphorylates BAD (Bcl2-antagonist of cell death) at S75. 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 (ASK1 and MST2), proliferation and angiogenesis (RB1). Can protect cells from apoptosis also by translocating to the mitochondria where it binds BCL2 and displaces BAD. Restricts caspase activation in response to selected stimuli, notably Fas stimulation, pathogen-mediated macrophage apoptosis, and erythroid differentiation.
Protein type: Protein kinase, TKL; Oncoprotein; Protein kinase, Ser/Thr (non-receptor); Kinase, protein; EC 2.7.11.1; TKL group; RAF family
Chromosomal Location of Human Ortholog: 3p25
Cellular Component: Golgi apparatus; mitochondrial outer membrane; cytoplasm; plasma membrane; pseudopodium; cytosol; nucleus
Molecular Function: small GTPase binding; protein serine/threonine kinase activity; identical protein binding; protein binding; MAP kinase kinase kinase activity; protein heterodimerization activity; mitogen-activated protein kinase kinase binding; metal ion binding; ATP binding; protein kinase activity
Biological Process: axon guidance; wound healing; activation of MAPKK activity; nerve growth factor receptor signaling pathway; somatic stem cell maintenance; apoptosis; heart development; intermediate filament cytoskeleton organization and biogenesis; negative regulation of caspase activity; signal transduction; protein amino acid phosphorylation; regulation of apoptosis; negative regulation of cell proliferation; synaptic transmission; small GTPase mediated signal transduction; regulation of cell differentiation; transmembrane transport; epidermal growth factor receptor signaling pathway; platelet activation; fibroblast growth factor receptor signaling pathway; adenylate cyclase activation; MAPKKK cascade; positive regulation of peptidyl-serine phosphorylation; cell proliferation; Ras protein signal transduction; response to hypoxia; insulin receptor signaling pathway; regulation of Rho protein signal transduction; innate immune response; blood coagulation; vascular endothelial growth factor receptor signaling pathway; negative regulation of protein complex assembly; negative regulation of apoptosis
Disease: Cardiomyopathy, Dilated, 1nn; Noonan Syndrome 5; Leopard Syndrome 2
Research Articles on RAF1
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Product Notes
The RAF1 raf1 (Catalog #AAA125994) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Phospho-RAF1-S621 Polyclonal Antibody reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's RAF1-S621 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunofluorescence (IF). WB: 1:500 - 1:2000 IF: 1:50 - 1:200. Researchers should empirically determine the suitability of the RAF1 raf1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "RAF1-S621, Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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