About

Contact Us

Email |
RSS

Protocols Online
Current and New Protocols for Life Science Research

You are here: Home » Recipes » PMSF

PMSF
Last update on July 13, 2012
under Recipes

Contents [hide]
Introduction
1. Introduction
In biochemistry, PMSF (phenylmethanesulfonylfluoride or phenylmethylsulfonyl fluoride)
2. Preparation of PMSF (10 mM), 10 ml
is a serine protease inhibitor commonly used in the preparation of cell lysates. PMSF
3. Recommended Suppliers
does not inhibit all serine proteases. It is rapidly degraded in water and stock solutions
are usually made up in anhydrous ethanol, isopropanol, corn oil, or DMSO. Proteolytic
inhibition occurs when a concentration between 0.1 – 1 mM PMSF is used. The half-life is short in aqueous solutions (110 min at
pH=7 and 35 min at pH=8).

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. Since PMSF binds covalently to the enzyme at the active serine residue, the complex can be viewed by X-ray
crystallography; it can therefore be used as a chemical label to identify an essential active site SER in an enzyme.

Enzyme (active) Ser-O-H + F-SO2CH2C6H5 ->  EnzymeSer-O-SO2CH2C6H5 + HF

Serine protease + PMSF -> Irreversible enzyme-PMS complex + HF

PMSF is a cytotoxic chemical that should only be handled inside a fume hood; the LD50 for this compound is less than 500 mg/kg.

Preparation of PMSF (10 mM), 10 ml

1. Weigh out  17.4 mg PMSF
2. Add isopropanol to 10ml and dissolve.
3. No need to filter sterilize.
4. Aliquot and store at –20°C.

PMSF is unstable in aqueous solutions with a half-life of approximately 30 minutes. Add solution imm

Recommended Suppliers
PMSF: Sigma (P-7626)

Share this:
Related

One-Dimensional SDS Gel Electrophoresis of Phosphate buffered saline DAB-Peroxidase Substrate Solution (Brown)
Proteins (western blotting) April 1, 2010 July 31, 2012
April 11, 2010 In “Recipes” In “Immunohistochemistry”
In “Electrophoresis”

Subscribe

Keep with with the latest news!

E-mail Submit

Related Posts:

Methyl Green Counterstain Protocol

Antibiotics Used in Molecular Biology
M9 medium
Polylysine-coated tissue culture surfaces
Tris Solution

Ampicillin
An Introduction to Mordants

Categories
EssentialsToggle
HistologyToggle
Molecular biologyToggle
Protein ScienceToggle
RecipesToggle

Latest
Popular
Comments
Tags

Methyl Green Counterstain Protocol December 19, 2018

DAB-Peroxidase Substrate Solution (Brown) July 31, 2012
Avidin-Biotin Blocking Solution July 24, 2012
SDS Antigen Retrieval Solution July 24, 2012
Tris Buffer Antigen Retrieval Protocol July 24, 2012

Protocols Online © 2016. All Rights Reserved.