Raf1 RBD Protein, Human, Recombinant, Biotinylated

Ras Binding Domain (RBD) of Human Raf-1 Proto-Oncogene, Serine/Threonine Kinase (RAF1), (UniProt # P04049, amino acids 50-140) with the N-terminal AviTag™ peptide and His-tag is produced in E. coli cells and site-specifically biotinylated at AviTag with BirA enzyme (https://amidbiosciences.com/collections/proteins/products/bira-biotin-ligase).

The Raf1 RBD protein binds specifically to GTP-bound Ras proteins. Ras proteins, like other small GTPases, regulate molecular events by cycling between an inactive GDP-bound form and an active GTP-bound form. In its active (GTP-bound) state, Ras binds specifically to the Ras-binding domain (RBD) of Raf1 to control downstream signaling cascades. The affinity between RBD and GDP-bound Ras is three orders of magnitude lower (1). Therefore, the RBD protein can be used to specifically precipitate active, GTP-bound Ras as well as to specifically block the activity of Ras in vitro and in vivo.

Applications

• Measurement of the GTP/GDP bound ratio of Ras in vitro.
• Quantitation of GTP-bound Ras from tissue and tissue culture cell lysates.
• RAS/RAF binding studies.
• Inhibitor screening for RAS/RAF interaction

MW: ~13.4 kDa
Tag(s): The N-terminal 6XHis and AviTag™
Amino Acids: 50-140 of Raf1, UniProt # P04049

Catalog # RAF1-B-301

Storage buffer: 20 mM HEPES, pH 7.5, 150 mM NaCl, 5 mM 2-mercaptoethanol, 1 mM MgCl₂, and 10% glycerol
Purity: >90% by Coomassie staining 
Storage is recommended at -80°C for longer periods of time. 

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This product is for laboratory research use only.

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References
de Rooij, J., Bos, J. Minimal Ras-binding domain of Raf1 can be used as an activation-specific probe for Ras. Oncogene 14, 623–625 (1997). https://doi.org/10.1038/sj.onc.1201005