General description

HRV-3C protease from human rhinovirus type 14 is a protease that specifically cleaves within an eight-residue recognition sequence.This sequence is: Leu-Glu-Val-Leu-Phe-Gln-Gly-Pro.Proteolytic cleavage occurs between the Gln and Gly residues. The HRV-3C protease is useful for cleaving recombinant proteins that are expressed as fusion proteins with this sequence between the carrier domain and the protein of interest.This biotinylated HRV-3C protease is intended for on-column cleavage of fusion proteins with an HRV-3C cleavage site. It specifically cleaves the protein of interest from a column-bound fusion protein, leaving the fusion domain or tag bound to the affinity column (e.g. Ni-NTA column) and eluting only the protein of interest. This method is advantageous over post-elution cleavage for several reasons: •It eliminates most impurities normally associated with purification on Ni-chelating columns. •It allows gentler elution conditions, with added flexibility in the elution buffer composition. This can mitigate protein aggregation and inactivation. After cleavage, the protease can be removed with any avidin-conjugated or streptavidin-conjugated beads. This product has been enzymatically biotinylated with no effect on its proteolytic activity. It has no additional protein purification tags. The product is supplied in aqueous buffer (0.8–1.2 mg/mL) with 20 mM Trizma^®-HCl, pH 8.0, 200 mM NaCl, 1 mM TCEP, and 50% (v/v) glycerol.

Legal Information

Trizma is a registered trademark of Sigma-Aldrich Co. LLC

Preparation Note

The product is supplied in aqueous buffer (0.8–1.2 mg/mL) containing 20 mM Trizma^®-HCl, pH 8.0, 200 mM NaCl, 1 mM TCEP, and 50% (v/v) glycerol.