RAF1 Antibody

Purified Mouse Monoclonal Antibody

Black line: Control Antigen (100 ng);
Purple line: Antigen(10ng);
Blue line: Antigen (50 ng);
Red line: Antigen (100 ng);
Figure 1: Western blot analysis using RAF1 mAb against human RAF1 (AA: 198-407) recombinant protein. (Expected MW is 73 kDa)
Figure 2: Western blot analysis using RAF1 mouse mAb against Hela (1), A431 (2), Cos7 (3) and C6 (4) cell lysate.
Figure 3: Immunohistochemical analysis of paraffin-embedded liver cancer tissues using RAF1 mouse mAb with DAB staining.
Figure 4: Flow cytometric analysis of Hela cells using RAF1 mouse mAb (green) and negative control (red).

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实验流程

Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC, FC, E
Primary Accession	P04049
Reactivity	Human
Host	Mouse
Clonality	Monoclonal
Clone Names	4G4
Isotype	IgG1
Calculated MW	73052 Da
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.
Immunogen	Purified recombinant fragment of human RAF1 expressed in E. Coli.
Formulation	Purified antibody in PBS with 0.05% sodium azide

Additional Information
Gene ID	5894
Other Names	RAF proto-oncogene serine/threonine-protein kinase, 2.7.11.1, Proto-oncogene c-RAF, cRaf, Raf-1, RAF1, RAF
Dilution	WB~~1/500 - 1/2000 IHC~~1/200 - 1/1000 FC~~1/200 - 1/400 E~~1/10000
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	RAF1 Antibody is for research use only and not for use in diagnostic or therapeutic procedures.

Protein Information
Name	RAF1 (HGNC:9829)
Synonyms	RAF
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.

Research Areas

For Research Use Only. Not For Use In Diagnostic Procedures.

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol