Turbo3C Protease, recombinant, Solution (HRV3C Protease)

HRV3C Protease (Human rhinovirus 3C Protease) is a cysteine protease which recognizes the cleavage site; Leu-Glu-Val-Leu-Phe-Gln↓Gly-Pro.
HRV3C Protease can be used for removal of tag from tag fusion protein which has the cleavage site of HRV3C Protease. Turbo3C Protease is HRV3C Protease with His tag and GST tag. Therefore Turbo3C Protease can be removed by either Ni chelating resin or GSH (Glutathione)
resin. Turbo3C Protease has significant activity at 4 °C, so it is difficult for other proteases to show activity and the target protein may be obtained with the stable form.

Source: E. coli expressed Turbo3C protease (human rhinovirus 3C protease)
Storage buffer: 25 mmol/L Tris-HCl (pH 8.0), 50 mmol/L NaCl, 1 mmol/L TCEP-HCl and 50 % Glycerol.
Activity: Indicated on each label.
Unit definition: 1 unit is the amount of enzyme which cleaves 95+ % of 100 μg of target protein at 4 °C for 16 hours.
Protein: Indicated on each label.
Molecular Weight: approximately 47,000
Cleavage site: Leu-Glu-Val-Leu-Phe-Gln↓Gly-Pro

Usage:

1. Cleavage condition
   1 unit of Turbo3C Protease cleaves 50~400 μg of the target protein in most cases. The efficiency of cleavage may vary with the sequences around the cleavage site and the conformation and the solubility of target protein. It is recommended to test using 1 unit of Turbo3C Protease to 100 μg of target protein in a suitable buffer for the target protein at 4 °C overnight. The final concentration of the target protein is 1~2 mg/mL. The optimum ratio should be determined after examining these results.
   Turbo3C Protease can be used at a higher ratio (1 unit: 10 μg) or for longer reaction time (over a few days) at higher temperature (37 °C) in order to get high cleavage efficiency because of its high specificity.
   Example of the reaction buffer:
   25 mmol/L Tris-HCl (pH 8.0), 150 mmol/L NaCl and 14 mmol/L 2-Mercaptoethanol.
   Note: Reaction buffer should be compatible with the downstream processes.
   For example, when Ni chelating resin will be used to remove the His tagged protein, minimum amount of EDTA or DTT should be used.


2. Removal of Turbo3C Protease
   After cleavage of the target protein, apply the reaction mixture of target protein and Turbo3C Protease to affinity columns if the reaction buffer is suitable for the affinity columns. Turbo3C Protease can be easily removed by affinity chromatography on a Ni chelating resin (for His tagged protein) or GSH resin (for GST tagged protein).