The configuration method and points for attention of PMSF

PMSF is a serine protease and acetylcholinesterase inhibitor. It inhibits proteases such as chymotrypsin, trypsin, papain, and mercapsin such as thrombin. PMSF inhibits the active site of sulfonated serine residues of serine proteases.

1.PMSF (phenylmethyl sulfonyl fluoride) dissolved in isopropanol, can not be dissolved in water. -20 ℃ preservation will also have crystallization precipitation, this is normal, until slightly higher temperature can be mixed.
2.PMSF severely damages respiratory mucosa, eyes and skin, and is fatal if inhaled, swallowed, or absorbed through the skin. Be sure to pay attention to protection, gloves and masks.
3. Once the eye or skin contact with PMSF, should be rinsed with a large amount of water immediately. Any clothing contaminated with PMSF should be discarded.
4.PMSF is unstable in aqueous solution. It should be added to the pyrolysis buffer from the storage solution before use.
5.The activity loss rate of 5.PMSF in aqueous solution increases with the increase of pH value, and the inactivation rate at 25 ℃ is higher than 4 ℃. When pH value is 8.0, the half-life of 20 μ mmol/l PMSF aqueous solution is about 85 min. This indicates that the PMSF solution can be safely discarded when it is adjusted to alkaline (pH > 8.6) and stored at room temperature for several hours.

Preparation method of PMSF:
The PMSF (phenylmethyl sulfonyl fluoride) dissolved in 174mg is in a sufficient amount of isopropanol, with a constant volume of 10 ml and a small fraction stored at -20 ℃. The final concentration is generally 1 mmol / L when used.

Introduction to use:
1) inhibition of serine protease (such as trypsin, chymotrypsin, thrombin) and mercaptoprotease (e.g. papain);
2) 10mg/ml was dissolved in isopropanol (or anhydrous ethanol);
3) it can be stored for one year at room temperature, generally at -20 degrees or 4 degrees.
4) working concentration: 17~174ug/ml (0.1~1mM), the concentration of storage solution was 100 or 200 mm;
5) unstable in the aqueous solution, it will degrade in half in 30 minutes. Fresh PMSF, samples must be added to each separation and purification step for more than 1 hour. See water decomposition, add before use

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