Sequence analysis of pure recombinant chymotrypsin how to dissolve, how to cut conditions - Database & Sql Blog Articles

Sequence analysis of pure recombinant chymotrypsin-related problems

Recommended method

Dissolved in 1 mM HCl or 50 mM HAc according to 1-10 mg/ml; in protein digestion, the amount of enzyme: target protein (mass ratio) = 1:20-1:100, temperature 30 °C, 50 mM Tris-HCl pH 7. 8 or 100 mM NH4HCO3, 5 mM CaCl2. When the temperature exceeds 37 ° C, the enzyme will self-cut.

store

The lyophilized powder is stored at 2-8 ° C and sealed against moisture. After dissolving, store at 2-8 °C, use within 2 days, or store at -20 °C after dispensing, and stabilize within 24 months. Avoid repeated freezing and thawing, and avoid avoiding long-term storage at room temperature.

1) How to dissolve chymotrypsin freeze-dried powder? Dissolved using 1 mM HCl or 50 mM HAc to a final concentration of 1-10 mg/ml.
2) What is the ratio of enzyme to protein when digested? According to the mass ratio, the amount of enzyme: the target protein (mass ratio) = 1: 20-1: 100, or according to the actual experimental results.
3) What is the optimum temperature, pH and buffer? The temperature was 30 ° C, pH 7.8, 40 mM Tris-HCl and 5 mM CaCl 2 .
4) The effect of excessive temperature on the enzyme? When the temperature exceeds 37 ° C, the enzyme will self-cut.