官方网址：http://www.sinobiological.cn/Protease-Tools-HRV-3C-Protease-g-1646.html
 
Content
    
        
            
2000 U
            
Recombinant HRV 3C Protease ( in 50 mM Tris, 150 mM NaCl, 1 mM EDTA, 50% Glycerol, pH 8.0 )
        
        
            
100 μg
            
Cleavage Control Protein ( lyophilized from sterile PBS , pH 7.4 )
        
        
            
5 ml
            
10X HRV 3C Cleavage Buffer ( 1.5 M NaCl , 0.5 M Tris-HCl , pH 7.5 )
        
    
 Description
HRV 3C Protease encoded by human rhinovirus 14 is a highly purified recombinant cysteine protease with a His-tag. Recombinant HRV 3C Protease is a ~20KDa single-chain protein containing approximately 189 amino acids with calculated pI 8.46. HRV 3C protease folds into two anti-parallel six-stranded β-barrels and the site cleft is located at the junction of the two β-barrels domains. The enzyme requires neither metal nor cofactors for activity. It has been demonstrated that the enzyme exhibits highest activity around neutral pH at temperature ranging from 22 to 37℃, even retaining robust activity at 4℃. Thus, cleavage can be performed at low temperature to enhance the stability of the target protein. The catalytic activity is insensitive to organic solvents (up to 10%); however, it can be strongly stimulated by high concentration of anions such as sulfate.
    
        
            
Specificity:
            
The enzyme recognizes the cleavage site: Leu-Glu-Val-Leu-Phe-Gln-↓-Gly-Pro
            
SDS-PAGE:
            
        
        
            
Molecular Weight:
            
~22KDa on SDS-PAGE
        
        
            
储存方法:
            
Store HRV 3C Protease at –20°C. Store HRV 3C Cleavage Control Protein and Protease Cleavage Buffer at –20°C or 4°C.
        
        
            
纯度:
            
98% by SDS-PAGE.
        
        
            
Quality Control:
            
The purity of each lot is determined by SDS-PAGE. And the activity is ensured by cleavage test with a recombinant fusion protein for each lot. The solution of HRV 3C protease is filtered through 0.22μm sterile filter before package.
        
    
 Application
The high specificity of HRV 3C protease makes it an ideal tool for cleaving fusion proteins at definite cleavage sites. The fusion protein can be purified and cleaved by HRV 3C to obtain the target protein. The recombinant HRV 3C protease is easily removed by IMAC Ni-charged resin.
 Activity definition
One unit of HRV 3C Protrease is defined as the amount of enzyme that will cleave >95% of 100μg HRV 3C cleavage control protein in 150mM NaCl, 50mM Tris-HCl pH 7.5, at 4℃ for 16h.
 User Protocol
Starting conditions
    
        
            
Temperature:
            
4℃
        
        
            
Incubation time:
            
16 hours or overnight