Member of the Rho family of GTPases1 within the superfamily of Ras-like GTPases. Similar to most GTPases, Rac1 cycles between GDP- and GTP-bound states; the GTP-bound form engages a set of diverse effectors to regulate cell processes. In particular, Rac1 promotes lamellipodia and membrane ruffles necessary for phagocytosis and cell migration. Nucleotide cycling is regulated by guanine nucleotide exchange factors (GEFs), guanine dissociation inhibitors (GDIs) and GTPase activating proteins (GAPs).
Protein is > 99% pure as determined by densitometry of the final sample submitted to SDS-PAGE and stained with Coomassie blue (Fig. 1). Rac1 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 1-189 of full-length, human Rac1. The terminal cysteine, which would normally be posttranslationally isoprenylated in eukaryotic cells, has been mutated to serine (C189S).
- Hodge RG, et al., Regulating Rho GTPases and their regulators. Nat Rev Mol Cell Biol 17, 496-510 (2016).
Species: Homo sapiens.
Sequence: Residues 1-189 of Rac1. The terminal cysteine is mutated to serine (C189S) and the N-terminus retains an Ala-Gly dipeptide remaining from the affinity tag.
Calculated molecular weight: 21,222 g/mol.
Experimental molecular weight: NA.
Extension coefficient: 22,920 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.