Raf-1 RBD Protein, GST, 300 µg

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.

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.

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.

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.

Glutathione-agarose

300 μg of Raf-1 RBD, GST in 150 μL in PBS, pH 7.4, 0.5mM PMSF, and 20% glycerol.

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

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