What is phenylmethylsulfonyl fluoride used for?

In biochemistry, phenylmethylsulfonyl fluoride (PMSF) is a serine protease inhibitor (serine hydrolase inactivator) commonly used in the preparation of cell lysates. PMSF does not inactivate all serine proteases.

How does PMSF inhibit trypsin?

Thermo Scientific PMSF is a protease inhibitor that reacts with serine residues to inhibit trypsin, chymotrypsin, thrombin and papain. In addition to inhibiting serine proteases, PMSF will also inhibit cysteine proteases, like papain (reversible by DTT treatment), and mammalian acetylcholinesterase.

What is PMSF solution?

Alternate Names: PMSF Solution is also known as Phenylmethylsulfonyl fluoride solution. Application: PMSF Solution is a nonspecific, irreversible inhibitor of serine proteases and other enzymes, including arylsulfatase A and trypsin. CAS Number: 329-98-6. Molecular Weight: 174.19.

Why is PMSF in lysis buffer?

PMSF is unstable in water and should be added to lysis buffers or other aqueous solutions just prior to use. Typically, a final concentration of 1 mM provides sufficient protease protection. Once in solution, PMSF can be stored at -20ºC for up to 3 months, protected from light.

How toxic is PMSF?

Phenylmethanesulfonyl fluoride (PMSF) is an acutely toxic and corrosive compound. Toxic if swallowed; has a lethal dose of 50% of the population (LD-50 oral, mouse) of 200 mg/kg. It causes severe eye damage, including potential blindness. Causes severe skin burns, irritation, inflammation, and/or blistering.

Is PMSF reversible?

Inhibition of the cysteine proteases by PMSF or AEBSF can be reversed by treatment with a sulfhydryl reagent such as DTT, but inhibition of serine proteases is irreversible.

Is PMSF irreversible?

It is an irreversible serine protease inhibitor, and is used at the same concentration as PMSF, but is nontoxic and stable in aqueous buffer solutions.

How long does PMSF last 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 does lysis buffer work?

Lysis buffers break the cell membrane by changing the pH. Detergents can also be added to cell lysis buffers to solubilize the membrane proteins and to rupture the cell membrane to release its contents. Chemical lysis can be classified as alkaline lysis and detergent lysis.

Why is PMSF toxic?

Phenylmethanesulfonyl fluoride (PMSF) is an acutely toxic and corrosive compound. PMSF hydrolyzes upon exposure to water liberating a toxic, acidic gas which in contact with metal surfaces can generate flammable and/or explosive hydrogen gas. Forms hydrofluoric acid upon exposure to water, which is extremely toxic.

What is PMSF and why is it added to your homogenization buffer?

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

Is PMSF toxic?

Phenylmethanesulfonyl fluoride (PMSF) is an acutely toxic and corrosive compound. Toxic if swallowed; has a lethal dose of 50% of the population (LD-50 oral, mouse) of 200 mg/kg. It causes severe eye damage, including potential blindness. Forms hydrofluoric acid upon exposure to water, which is extremely toxic.