19 Need-to-Know Facts About PMSF Inhibitors:
PMSF or phenylmethylsulfonyl fluoride is a serine protease inhibitor. It targets serine proteases, including trypsin, chymotrypsin, thrombin, and thiol proteases such as papain. Typically its effect is irreversible with the exception of DTT treatment.
PMSF is commonly used in protein solubilization in order to deactivate proteins of interest after cell lysis.
Molecular Formula: C7H7FO2S
Molecular Weight: 174.19
Inhibition: Proteolytic inhibition occurs when a concentration between 0.1 – 1 mM of PMSF is used.
CAS #: 329-98-6
Synonyms: phenylmethane sulfonyl fluoride or phenylmethylsulfonyl fluoride
Melting Point: 91-95°C
What happens to PMSF in water?
PMSF has limited solubility and is highly unstable in aqueous solutions. It should be added to buffers immediately before use. The half-life of an aqueous solution of PMSF is around 30 minutes at pH 8.
Is PMSF hazardous?
PMSF is a cytotoxic chemical which should be handled only inside a fume hood or certified Biological Safety Cabinet. Lethal dose is less than 500 mg/kg. RTECS: XT8040000, UN2928, Packaging group: II.
Is there a non-toxic alternative?
AEBSF, (4-[2-aminoethyl]-benzenesulfonylfluoride, hydrochlorid or Pefabloc) is a non-toxic alternative. 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 do I make a PMSF solution?
Stock solutions are usually made up in anhydrous ethanol, isopropanol, or DMSO. PMSF is also soluble in ether, benzene, (anhydrous 2-propanol (200 mM), anhydrous methanol (200 mM), and anhydrous ethanol (200 mM).
Effective in most protein solutions at 0.1 to 1 mM final concentration. Dissolve PMSF in isopropanol at 17.4 mg/mL (100 mM) and store in aliquots at -20°C.
1. Weigh out 17.4 mg PMSF.
2. Add isopropanol to 10ml and dissolve.
3. No need to filter or sterilize.
4. Aliquot and store at –20°C.
Does PMSF require special handling?
PMSF is cytotoxic and hygroscopic, and should be protected from light.
What happens to PMSF if you increase pH and temperature?
The rate of inactivation increases with increasing pH and is faster at 25°C than at 4°C.
What is the half-life of PMSF?
The half-life of a 20 mM aqueous solution of PMSF is ~35 min at pH 8.0.
Elaborate on the PMSF GHS classification.
GHS-US, H301 classification acute toxicity 3 (oral), skin H314, Corrosive 1B, Eye Damage H318 1.
What is the PMSF binding site?
PMSF binds specifically to the active site serine residue in a serine protease, but does not bind to any other serine residues in the protein.
Can PMSF be utilized in X-ray crystallography?
PMSF binds covalently to the enzyme at the active serine residue of which can be viewed by X-ray crystallography; therefore, PMSF can be used as a chemical label to identify an essential active site serine in an enzyme.
What are the theoretical levels for Carbon & Hydrogen?
Carbon: 48.26 and Hydrogen: 4.05.
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