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HRV3C Protease: Turbo3C

Human rhinovirus 3C protease (HRV3C Protease) is a cysteine protease that recognizes the cleavage site of Leu-Glu-Val-Leu-Phe-Gln/Gly-Pro, commonly referred to as the PreScission Site. It cleaves between Gln and Gly. The recombinant Turbo3C Protease is a restriction grade HRV3C protease that has robust activity at 4°C and high specific activity. Turbo3C Protease is a 47 kDa protein with both GST and His tags so it can be removed by either Ni chelating or Glutathione (GSH) resin.

Features

  • Affordable ultra-pure HRV3C Protease
  • Activity over a broad temperature (4°C to 37°C) and pH (6.5 to 8.5) range
  • Both GST and His tags to facilitate its removal from the digested protein sample

Activity and Specificity

The activity of Turbo3C Protease is tested using a control target protein. 1 ug of Turbo3C Protease has at least 1 unit activity conventionally used by other suppliers (1 unit of HRV3C protease cleaves >95% of 100 ug of control target protein at 4°C for 16 hours). No non-specific activity has been observed under the same condition with Turbo3C Protease to control target protein ratio of 1:10. Prolonged incubation (several days) under the same condition does not show any non-specific cleavage.

Figure 1
A 68 kDa GST-fusion protein (C) at 1 mg/ml is incubated with Turbo3C Protease (*) at a ratio of (1) 1:50, (2) 1:100, (3) 1:200, (4) 1:400 (w/w) in a buffer of 25 mM Tris-HCl, pH8.0, 150 mM NaCl, 14 mM 2-mercaptoethanol at 4°C for 16 hours. The cleaved products are 42 kDa and 26 kDa

Formulation and Storage

2 mg/ml in 50 mM Tris-HCl, pH8.0, 150 mM NaCl, 1 mM EDTA, 1 mM TCEP, 50% glycerol. Store at -20°C. Shipped in gel packs.

Source

Purified from E. coli expressing the Turbo3C protease gene.

Cleavage Condition

It is recommended to use Turbo3C Protease at a protease-to-target protein ratio of 1:100 (w/w) or 1 unit of Turbo3C Protease to 100 ug of target protein in a buffer suitable for the target protein at 4°C overnight, with the target protein concentration at 1-2 mg/ml. In most cases, target proteins are completely cleaved with a protease-to-target protein ratio of 1:50 to 1:400 or 1 unit Turbo3C Protease to 50-400 ug of target protein (as shown in Figure 1). The efficiency of cleavage may vary due to the sequences around the cleavage site, the conformation and the solubility of the target protein. Due to its high specificity, more Turbo3C Protease (at 1:10 ratio) or longer cleavage time (over a weekend) at higher temperature (37°C) can be used to achieve high cleavage efficiency without non-specific cleavage of target proteins.

Removal of Turbo3C Protease after Cleavage

The Turbo3C Protease contains both GST and His tags. After cleavage of the target protein, Turbo3C Protease can be easily removed along with the tags from the cleavage reaction by affinity chromatography on a Ni-chelating resin for His-tagged target protein or GSH resin for GST-tagged target protein.

Technical Information

Turbo3C Procedure (PDF)

Ordering Information

Product Cat.No. Storage Quantity Price(US$)  
Turbo3C (HRV3C Protease) 2 mg/ml NU0101S -20°C 1,000 units (1 mg) 250.00 Buy
Turbo3C (HRV3C Protease) 2 mg/ml NU0101M -20°C 10,000 units (10 mg) 2,100.00 Buy