9014-74-8 Usage
Description
Enterokinase is a highly specific serine protease enzyme found in the small intestine, which converts trypsinogen into trypsin. It is used for the removal of the FLAG peptide from N-terminal and Met-N-terminal fusion proteins, but does not remove the C-terminal FLAG.
Uses
Used in Biochemistry and Molecular Biology:
Enterokinase is used as a cleavage agent for removing the FLAG peptide from N-terminal and Met-N-terminal fusion proteins. This is important for the purification and analysis of recombinant proteins in research and biotechnology applications.
Used in Protein Purification:
Enterokinase is used as a specific protease for the selective cleavage of fusion proteins, allowing for the purification of the target protein of interest. This is particularly useful in the production of recombinant proteins for therapeutic or research purposes.
Typical conditions for fusion protein cleavage involve adjusting the concentration of the fusion protein to 1.5 mg/ml and a pH between 7.0-8.0 with 500 mM Tris-HCl, pH 8.0, 2.0 mM CaCl2, and 1% Tween 20. Enterokinase is then added to the fusion protein solution at a ratio of approximately 0.02 units per 1 mg fusion protein and mixed. The reaction mixture is incubated at approximately 25°C for 16 hours to allow for efficient cleavage of the fusion protein.
Biochem/physiol Actions
Enterokinase is a membrane bound serine protease that specifically and rapidly converts trypsinogen to trypsin, thereby, triggering the conversion of other zymogens to active enzymes. It has a molecular mass of approximately 150 kDa. The enzyme is a heterodimer consisting of 35-47 kDa subunits. The light and the heavy chains are linked by two disulfide bridges. It is a glycoprotein containing 35% carbohydrate. The polypeptide chain of trypsinogen is hydrolyzed only after an -(Asp)4-Lys- sequence. The enzyme is inhibited by soybean trypsin inhibitor. Enterokinase is typically used in protein modification and amino acid sequence determination.
Check Digit Verification of cas no
The CAS Registry Mumber 9014-74-8 includes 7 digits separated into 3 groups by hyphens. The first part of the number,starting from the left, has 4 digits, 9,0,1 and 4 respectively; the second part has 2 digits, 7 and 4 respectively.
Calculate Digit Verification of CAS Registry Number 9014-74:
(6*9)+(5*0)+(4*1)+(3*4)+(2*7)+(1*4)=88
88 % 10 = 8
So 9014-74-8 is a valid CAS Registry Number.
