HRV 3C Protease is a human rhinovirus (HRV) 14 3C protease with a 6XHis tag at the C-terminus expressed and purified from Escherichia coli. HRV 3C Protease specifically recognizes an 8-residue peptide motif of LeuGluValLeuPheGln/GlyPro and cleaves between Gln and Gly residues of the recognition sequence. HRV 3C Protease appears at the molecular weight of approximately 24 kD on the reduced SDS-PAGE gel. The efficiency may vary with different fusion partners. Optimal conditions for cleavage of each fusion can be achieved through trials prior to the tag removal. After cleavage of the fusion protein, the polyhistidine tag can be utilized to remove the HRV 3C protease from the cleavage reaction by immobilized nickel affinity chromatography. Dialysis against the loading buffer may be necessary to remove EDTA in the cleavage reaction solution for nickel resin.
Applications:
Features:
Applications:
- HRV 3C Protease recognizes the 8-residue sequence and specifically cleaves at the site between glutamine and glycine: Leu-Glu-Val-Leu-Phe-Gln-↓-Gly-Pro. It is therefore usually used to remove fusion tags from proteins with an HRV 3C cleavage site.
Features:
- Purity >98% by Coomassie Blue Staining
- Specific activity 2000 U/mg or 1000 U/ml
- Endotoxin ≤ 0.1 EU/ml (0.0001 EU/ 1000 U)
- Unit Definition: One unit is defined as the amount of enzyme needed to cleave 100 μg of fusion protein in 16 hours to 90% completion at 4°C in a buffer of 50 mM Tris-HCl at pH 7.0 with 150 mM NaCl, 1 mM EDTA, and 1 mM dithiothreitol (DTT).
- Cleavage buffer 50 mM Tris-HCl at pH 7.0 with 150 mM NaCl, 1 mM EDTA, and 1 mM DTT.
- A 6XHis tag at C-terminus can be utilized for removing HRV 3C Protease from the cleavage reaction solution.
- Storage and Shipping: Stored in the buffer of 50 mM Tris-HCl at pH 8 with 150 mM NaCl, 5 mM EDTA, and 1 mM DTT. Store at -20°C upon arrival. Avoid repeated freeze-thaw cycles.