14-863
Raf-1 RBD Protein, GST, 300 µg
GST fusion-protein, corresponding to the human Ras Binding Domain (RBD, residues 1-149) of Raf-1, expressed in E. coli. with purity 50% at full length molecular weight 42 kDa. For use in Affinity Binding Assays
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About This Item
UNSPSC Code:
12352202
eCl@ss:
32160405
NACRES:
NA.41
Recommended Products
biological source
human
Quality Level
form
liquid
shelf life
1 yr
mol wt
Mw 42 kDa
manufacturer/tradename
Upstate®
concentration
>50%
technique(s)
affinity binding assay: suitable
solubility
water: soluble
NCBI accession no.
UniProt accession no.
shipped in
dry ice
storage temp.
−20°C
Gene Information
human ... RAF1(5894)
General description
Research Area: Cell signaling
The rapidly accelerated fibrosarcoma (Raf) family, consisting of three highly conserved cytosolic serine/threonine kinases (ARAF, BRAF, and RAF1 or CRAF) is characterized by three conserved regions such as CR1, CR2, and CR34. The RAS-binding domain (RBD) and the cysteine-rich domain (CRD) are found in the N-terminal CR1 region.
The rapidly accelerated fibrosarcoma (Raf) family, consisting of three highly conserved cytosolic serine/threonine kinases (ARAF, BRAF, and RAF1 or CRAF) is characterized by three conserved regions such as CR1, CR2, and CR34. The RAS-binding domain (RBD) and the cysteine-rich domain (CRD) are found in the N-terminal CR1 region.
Application
Research Category
Metabolism
Metabolism
Research Sub Category
Obesity
Metabolic Disorders
Obesity
Metabolic Disorders
Specifically binds to and precipitates Ras-GTP from cell lysate.Can be bound to glutathione coated plates prior to precipitating Ras-GTP.
Biochem/physiol Actions
c-Raf-1 acts as an effector of Ras and is a serine-threonine kinase that transmits incoming signals to the mitogen-activated protein kinase pathway. This pathway is associated with various cellular responses, including cell proliferation, survival, and differentiation. RBD directly binds to the Ras·GTP complex in vitro. Furthermore, bacterially expressed recombinant RBD assumes a stable folded conformation in vitro. The GST·RBD fusion protein, linked to Sepharose beads may be used to precipitate oncogenic Leu61-Ha-Ras, which is constitutively bound with GTP.
Quality
Affinity Precipitation of Activated Ras: Raf-1-RBD, GST can be captured on a glutathione coated plate and used to capture GTP activated Ras from HeLa cell lysate. Used 2 μg of Raf-1-RBD, GST per well to capture 25-100 μg of EGF stimulated HeLa cell lysate. Upstate also offers a Ras GTPase ELISA kit (17-424).
Note: Due to spontaneous deactivation of Ras, use lysates that have been stored at -70°C for less than 6 month.
Note: Due to spontaneous deactivation of Ras, use lysates that have been stored at -70°C for less than 6 month.
Physical form
300 μg of Raf-1 RBD, GST in 150 μL in PBS, pH 7.4, 0.5mM PMSF, and 20% glycerol.
Glutathione-agarose
Storage and Stability
1 year at -20°C from date of shipment.
Legal Information
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage Class Code
10 - Combustible liquids
WGK
WGK 2
Certificates of Analysis (COA)
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Point mutants of c-raf-1 RBD with elevated binding to v-Ha-Ras
Fridman M, et al.
The Journal of biological chemistry, 275(39), 30363-30371 (2000)
R Wynn et al.
Protein science : a publication of the Protein Society, 6(8), 1621-1626 (1997-08-01)
We have determined by X-ray crystallography the structures of several variants of staphylococcal nuclease with long flexible straight chain and equivalent length cyclic unnatural amino acid side chains embedded in the protein core. The terminal atoms in the straight side
Timothy H Tran et al.
Nature communications, 12(1), 1176-1176 (2021-02-21)
The first step of RAF activation involves binding to active RAS, resulting in the recruitment of RAF to the plasma membrane. To understand the molecular details of RAS-RAF interaction, we present crystal structures of wild-type and oncogenic mutants of KRAS
M Foschi et al.
The EMBO journal, 16(21), 6439-6451 (1997-11-14)
Endothelin-1 (ET-1) induces cell proliferation and differentiation through multiple G-protein-linked signaling systems, including p21ras activation. Whereas p21ras activation and desensitization by receptor tyrosine kinases have been extensively investigated, the kinetics of p21ras activation induced by engagement of G-protein-coupled receptors remains
Elizabeth M Terrell et al.
Molecular cell, 76(6), 872-884 (2019-10-14)
The Ras GTPases are frequently mutated in human cancer, and, although the Raf kinases are essential effectors of Ras signaling, the tumorigenic properties of specific Ras-Raf complexes are not well characterized. Here, we examine the ability of individual Ras and
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