how to make pmsf stock solution

How do you make a 100mm PMSF solution?

To prepare a 100 mM solution, add 17.4 mg of PMSF per milliliter of isopropanol. Store at −20°C. PMSF is inactivated in aqueous solutions. The rate of inactivation increases with increasing pH and is faster at 25°C than at 4°C.

In which solvent would you dissolve PMSF and why?

PMSF is considered a highly toxic cholinesterase inhibitor. This product is soluble at 0.2 M (200 mM) in isopropanol. When needed, sonication (or gentle heating, if absolutely necessary) may be applied to dissolve fully the PMSF.

What is PMSF dissolved in?

PMSF is soluble in organic solvents such as ethanol, DMSO, and dimethyl formamide (DMF), which should be purged with an inert gas. The solubility of PMSF in ethanol and DMSO is approximately 20 mg/ml and approximately 33 mg/ml in DMF. PMSF is sparingly soluble in aqueous buffers.

How long is PMSF stable in solution?

The half-life of a 20mM aqueous solution of PMSF is ~35 minutes at pH 8.0. This short half-life means that aqueous solutions of PMSF can be safely discarded after they have been rendered alkaline (pH >8.6) and stored for several hours at room temperature.

How do you handle PMSF?

Keep containers tightly closed. PMSF may corrode/etch glass; therefore do not store aqueous solutions of PMSF in glass containers. Store in a cool, dry, and well-ventilated area away from incompatible chemicals such as aqueous solutions, strong oxidizing agents, strong bases, and acids.

What is PMSF concentration?

between 0.1 - 1 mMThe effective concentration of PMSF is between 0.1 - 1 mM. The half-life is short in aqueous solutions (110 min at pH 7, 55 min at pH 7.5, and 35 min at pH 8, all at 25 °C). Stock solutions are usually made up in anhydrous ethanol, isopropanol, or corn oil and diluted immediately before use.

What is PMSF for in lysis buffer?

Product Description. Phenylmethanesulfonyl Fluoride (PMSF) is an inhibitor of serine proteases such as trypsin, chymotrypsin, thrombin, and papain. It is routinely added as a supplement to lysis buffers just prior to lysis, to prevent protease degradation.

Why is PMSF added to extraction buffer?

PMSF is a protease inhibitor that was added to prevent degradation of proteins of interest.

Is PMSF reversible?

Active Motif's Phenylmethylsulfonyl fluoride (PMSF) is a non-specific inhibitor of serine proteases and the cysteine protease papain (reversible by DTT treatment). PMSF does not inhibit metalloproteases, most cysteine proteases or aspartic proteases.