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Raf protein ras binding domain on GST beads

貨號 RF02-A 售價(jià)(元) 7498
規(guī)格 1 x 2 mg CAS號
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Product Uses Include:

Measurement of the GTP/GDP ratio of Ras in vitro.

Quantitation of GTP-Ras from tissue and tissue culture cell lysates.

Material:
       The Ras binding domain (RBD) of the c-Raf kinase protein (Raf) protein has been expressed as a GST-fusion protein in E. coli. This protein binds binds specifically to GTP-bound, and not GDP-bound, Ras proteins. The domain can therefore be used to specifically precipitate active, GTP-bound Ras as well as to specifically block the activity of Ras in vitro and in vivo.

       The protein is supplied in a glutathione agarose bound format and is shipped lyophilized. The beads are colored for ease of use. This product is used in our Ras activation assay Biochem Kit? (Cat. # BK008). The GST-tagged Raf-RBD protein can be released from the beads by incubation with 10 mM reduced glutathione.

 

       Figure 1. The brightly colored glutatione agarose beads in RF02 are easy to use.

Purity:
       Protein purity is determined by scanning densitometry of Coomassie Blue stained protein on a 12% SDS polyacrylamide gel. GST-Raf-RBD protein is >80% pure (see Figure 2).

 

        Figure 2: GST-Raf-RBD protein purity determination. A 20 μg sample of RF02 was separated by electrophoresis in a 12% SDS-PAGE system and stained with Coomassie Blue. The GST-Raf-RBD protein runs at approximately 35 kDa.

Biological Activity:
        Raf-RBD protein specifically recognizes and binds the active, GTP-bound, forms of Ras proteins. It has a much lower affinity for the inactive, GDP-bound, form Ras. When coupled to a colored glutathione sepharose matrix, the Raf-RBD protein beads become a convienent tool for assaying the activity of the Ras proteins. The quality control biological assay for Raf-RBD protein beads consists of a Ras protein pulldown frombovine brain extracts loaded with either GTPγS (Cat. # BS01) or GDP.

 

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