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

• UNSPSC Code: 12352202
• NACRES: NA.41
• eCl@ss: 32160405
• Form: liquid
• Biological source: human
• Mol wt: M_w 42 kDa

Eigenschaften

• biological source: human
• Quality Segment: 100
• form: liquid
• shelf life: 1 yr
• mol wt: M_w 42 kDa
• manufacturer/tradename: Upstate^®
• concentration: >50%
• technique(s): affinity binding assay: suitable
• solubility: water: soluble
• NCBI accession no.: NM_002880.2
• UniProt accession no.: P04049
• shipped in: dry ice
• storage temp.: −20°C
• Gene Information: human ... RAF1(5894)

Beschreibung

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.

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

Preparation Note

1 year at -20°C from date of shipment.

Analysis Note

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.

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.

Sicherheitshinweise

• Lagerklasse: 10 - Combustible liquids
• wgk: WGK 2