Cat. # RN05

Product Uses Include

• Ran GEF binding studies
• Inhibition of Ran GEFs in vitro
• Inhibition of Ran proteins in vivo by microinjection
  
  

Material
The dominant negative form of human Ran protein contains a threonine to asparagine substitution at residue 24. The common name for this mutant is Ran(T24N) or N24Ran. The asparagine substitution abolishes the protein's affinity for GTP and reduces its affinity for GDP. Hence, the Ran(T24N) is always in either a nucletiode free state or in its inactive, GDP-bound, state. Because of this, it binds strongly to Ran GEFs and it blocks wild type Ran from being activated by these GEFs.

The recombinant protein is approximately 35 kDa, consisting of the Ran protein plus a histidine tag in the amino-terminus. Protein concentration is determined by the Precision Red Advanced Protein Assay Reagent, Cat. # ADV02.

For other forms of Ran (wild type, constitutively active) as well as many other purified small G-proteins, see our main small G-protein product page.

Purity
Purity is determined by scanning densitometry of proteins on SDS-PAGE gels. His-Ran samples are >90% pure.

Figure 1: His-Ran(T24N) protein purity determination. A 10 µg sample of RN05 was separated by electrophoresis in a 12% SDS-PAGE system. The protein was stained with Coomassie Blue.