Product Introduction
|
Product Name |
Cat. No. |
Spec. |
|
Recombinant Protease (His-tag) |
G3410-1000U |
1000 U |
Product Description/Introduction
Recombinant HRV 3C Protease is a recombinant cysteine protease from human rhinovirus type 14 3C with His tag expressed in Escherichia coli. HRV 3C Protease cleaves with the recognition sequence L-E-V-L-F-Q-G-P (LeU-Glu-Val-Leu-Ph-Gln-Gly-Pro) or the core pentapeptide sequence L-F-Q-G-P (Leu-Ph-Gln-Gly-Pro) at low temperature between the Q (Gln) and G-P (Gly-Pro) residues. The high specificity and activity of the protease make it still active even at low temperature of 4℃.
Application: Remove MBP, GST, His or other tag proteins from fusion proteins by binding HRV 3C protease which contains His tag to the nickel column.
Source: Human Rhinovirus type 14 3C Protease, recombinant expressed in Escherichia coli, containing His tag.
Enzyme activity : It is defined as one active unit for cleaving more than 90% of 100 μg substrate (protein containing HRV 3C Protease cleavage site) within 16 h at 4℃, pH 7.5.
Inactivation or inhibition: HRV 3C Protease activity was inhibited by 100 mM ZnCl2, 4 mM AEBSF, and 100 μM Chymostatin.
Purity and concentration: Purity ≥95% by SDS-PAGE, 2 U/μL.
Recombinant HRV 3C Protease storage solution：50 mM Tris-HCl,150 mM NaCl,1 mM EDTA,1 mM DTT,50% (v/v) glycerol,pH 8.0.
10×HRV 3C Buffer：500 mM Tris-HCl,1.5 M NaCl,10 mM EDTA,10 mM DTT,pH 7.5.
Storage and Shipping Conditions
Ship with wet ice; Store at -20℃, valid for 12 months.
Product Components
|
Component Number |
Component |
G3410-1000U |
|
G3410-1 |
Recombinant HRV 3C Protease (His-tag) (2 U/μL) |
500 μL |
|
G3410-2 |
10×HRV 3C Buffer |
3×1 mL |
|
G3410-3 |
Control Protein |
100 μL |
|
Manual |
||
Assay Protocol/Procedures
1. Protease digestion system recommended：
|
Component |
Volume |
|
10×HRV 3C Buffer |
5 μL |
|
Target Proteins |
200 μg |
|
Recombinant HRV 3C Protease (His-tag) (2U/μL) |
1 μL |
|
ddH2O |
To 50 μL |
2. Place at 4℃ overnight or for 16 h;
3. Remove 10 μL of enzyme digestion product for SDS-PAGE electrophoresis to evaluate the amount of enzyme digestion and reaction conditions.
Note
1. Enzyme products should be placed in an ice box or ice bath when used and stored separately at -20℃ immediately after use;
2. For your safety and health, please wear safety glasses, gloves, or protective clothing.
Examples of enzyme digestion applications
Figure 1: Effect of HRV 3C Protease digestion of control substrate protein(A+B fusion protein, which contains HRV 3C Protease Enzyme cutting site) (The restriction conditions are 30℃, pH 7.5, 0 min, 10 min, 20 min, 30 min, 1 h, 2 h, respectively, corresponding to Lane: 1-6, the mass ratio of added substrate to enzyme in the enzymatic digestion system is 10:1)
For Research Use Only!
