Member of family of Rho GTPases1 within the superfamily of Ras-like GTPases. Similar to most GTPases, RhoA cycles between GDP- and GTP-bound states; the GTP-bound form engages a set of diverse effectors to regulate cell processes. In particular, RhoA is associated with cytoskeleton regulation, mostly through the formation of actin stress fibers and actomyosin contractility. Nucleotide cycling is regulated by guanine nucleotide exchange factors (GEFs), guanine dissociation inhibitors (GDIs) and GTPase activating proteins (GAPs).
Protein is > 96 % pure as determined by densitometry of the final sample submitted to SDS-PAGE and stained with Coomassie blue (Fig. 1). RhoA was heterologously expressed in E. coli as a histidine-tagged protein and purified by a combination of affinity, ion exchange, and gel exclusion chromatographies. The tag was removed prior to final purification leaving a Gly-Ala dipeptide at the N-terminus followed by residues 1-190 of full-length, human RhoA. The terminal cysteine, which would normally be posttranslationally isoprenylated in eukaryotic cells, has been mutated to serine (C190S).
- Hodge RG, et al., Regulating Rho GTPases and their regulators. Nat Rev Mol Cell Biol 17, 496-510 (2016).
Species: Homo sapiens.
Sequence: Residues 1-190 of RhoA. The terminal cysteine is mutated to serine (C190S) and the N-terminus retains an Ala-Gly dipeptide remaining from the affinity tag.
Calculated molecular weight: 21,554 g/mol.
Experimental molecular weight: NA.
Extension coefficient: 18,450 M-1cm-1 (assuming all cysteines remain reduced).
Amount: 100 or 500 μg.
Concentration: 1 mg/ml (determined by absorbance at 280 nm).
Buffer: 25 mM Tris pH 7.5, 150 mM NaCl, 5% glycerol and 1mM DTT.
Form: Frozen liquid.
Storage and stability:
Storage: -80°C. After initial thaw, aliquot and store at -80°C until further use.
Stability: Stable for up to 12 months at -80°C and up to 2 days at 4°C. Repeated freeze-thaw cycles not recommended.
Notes: Please see accompanying sheet describing activity.
Precautions and disclaimer:
This product is for in vitro research only and not for drug, household, or other use in humans and animals.