Recombinant Raf1 (mutated Y340E + Y341E) protein (Active) is a Human Fragment protein, in the 306 to 648 aa range, expressed in Baculovirus infected Sf9, with >80% purity and suitable for FuncS, WB.
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Key facts
- Purity
- >80% SDS-PAGE
- Expression system
- Baculovirus infected Sf9 cells
- Tags
- GST tag N-Terminus
- Applications
- FuncS, WB
- Biologically active
- Yes
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application FuncS | Reactivity Reacts | Dilution info - | Notes - |
Application WB | Reactivity Reacts | Dilution info - | Notes - |
Target data
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.
Alternative names
RAF, RAF1, RAF proto-oncogene serine/threonine-protein kinase, Proto-oncogene c-RAF, Raf-1, cRaf
Recommended products
Recombinant Raf1 (mutated Y340E + Y341E) protein (Active) is a Human Fragment protein, in the 306 to 648 aa range, expressed in Baculovirus infected Sf9, with >80% purity and suitable for FuncS, WB.
Key facts
- Purity
- >80% SDS-PAGE
- Expression system
- Baculovirus infected Sf9 cells
- Tags
- GST tag N-Terminus
- Applications
- FuncS, WB
- Biologically active
- Yes
- Biological activity
- The specific activity of RAF1 was determined to be ~6,000 nmol/min/mg in a coupled assay as per antibody assay protocol.
- Accession
- P04049-1
- Animal free
- No
- Species
- Human
- Concentration
- Storage buffer
pH: 7.5
Constituents: 25% Glycerol (glycerin, glycerine), 0.87% Sodium chloride, 0.79% Tris HCl, 0.31% Glutathione, 0.00385% (R*,R*)-1,4-Dimercaptobutan-2,3-diol, 0.0029% EDTA, 0.0017% PMSF
Sequence info
Amino acid sequence
- Accession
- P04049
- Protein length
- Fragment
- Predicted molecular weight
- 63 kDa
- Amino acids
- 306 to 648
- Nature
- Recombinant
- Tags
- GST tag N-Terminus
Specifications
- Form
- Liquid
- Additional notes
Purity >80% as determined by SDS-PAGE and Coomassie blue staining. Affinity purified.
General info
- 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.
- Sequence similarities
Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. RAF subfamily.
- 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 SHOC2-MRAS-PP1c (SMP) complex consisting of SHOC2, GTP-bound M-Ras/MRAS and the catalytic subunit of protein phosphatase 1 (PPP1CA, PPP1CB or PPP1CC); this relieves inactivation and stimulates kinase activity (PubMed:35768504, PubMed:35831509, PubMed:35830882). 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 an activity decrease.
- Subcellular localisation
- Mitochondrion, Nucleus
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- -80°C
- Appropriate long-term storage conditions
- -80°C
- Aliquoting information
- Upon delivery aliquot
- Storage information
- Avoid freeze / thaw cycle
This product is an active protein and may elicit a biological response in vivo, handle with caution.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Raf1 also known as c-Raf or Raf-1 is a serine/threonine-protein kinase that plays an important role in cell division differentiation and survival. Its molecular weight is approximately 74 kDa. Raf1 is part of the MAPK/ERK signaling pathway and is ubiquitously expressed in many tissues including the brain heart and liver. As a component of the signaling cascade its activation translates extracellular signals into intracellular responses.
- Biological function summary
Raf1 regulates important cellular processes by activating downstream kinases in response to external stimuli. Raf1 forms a complex with other proteins such as Ras facilitating its role as an essential component of the MAPK/ERK pathway. Upon activation by Ras Raf1 phosphorylates and activates MEK1 and MEK2 which in turn activate the ERK1 and ERK2. This signaling axis is involved in controlling gene expression and cellular proliferation.
- Pathways
Raf1 is integrally involved in the MAPK/ERK pathway which is critical for transducing signals from growth factors and mitogens. It relates closely with proteins such as Ras MEK and ERK in this pathway. The pathway is important for regulating cellular responses to various stimuli and is particularly involved in processes such as cell cycle control and apoptosis.
- Associated diseases and disorders
Raf1 has associations with certain types of cancer and Noonan syndrome. In various cancers mutations or dysregulation of Raf1 or related components such as Ras can lead to uncontrolled cellular proliferation. In Noonan syndrome Raf1 mutations result in anomalies in the Ras/MAPK pathway which can impact normal development. Raf1's interactions in these pathways indicate its relevance as a potential target for therapeutic intervention.
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2 product images
Functional Studies - Recombinant Raf1 (mutated Y340E + Y341E) protein (Active) (ab62292)
ab62292 Kinase assay plot
Western blot - Recombinant Raf1 (mutated Y340E + Y341E) protein (Active) (ab62292)
Lane 1: Western blot - Recombinant Raf1 (mutated Y340E + Y341E) protein (Active) (ab62292)
Lane 1: RAF1
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