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Key Documents

14-863

Sigma-Aldrich

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

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.

Application

Research Category
Metabolism
Research Sub Category
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.

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

10 - Combustible liquids

wgk_germany

WGK 2


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Navštívit knihovnu dokumentů

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
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)
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
J de Rooij et al.
Oncogene, 14(5), 623-625 (1997-02-06)
Ras is a small GTPase that cycles between an inactive GDP-bound and an active GTP-bound form. A large variety of ligands that stimulate cell surface receptors induce the activation of Ras. Thus far, this activation could only be measured by
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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