Description
Pronase is a mixture of several nonspecific endo- and exoproteases that digest proteins down to single amino acids.
Abbr
Pronase, Native (Streptomyces griseus)
Source
Streptomyces griseus
Applications
Use Pronase to completely hydrolyze proteins in research applications. Pronase is used for the degradation of proteins during the isolation of DNA and RNA, such as in the extraction of phage DNA or the isolation of plasmid DNA. It is not necessary to let pronase self-digest prior to use. It is also used in histochemistry and cell culture for tissue dissociation in conjunction with collagenase and trypsin, and for the production of glycopeptides from purified glycoproteins.
Enzyme Commission Number
EC 3.4.24.4
Activity
~7.0 units/mg protein (at 40°C with casein as the substrate, pH 7.5, equivalent to approximately 1270 PU/mg or approximately 25 PUK/mg.)
Specificity
Pronase has a broad specificity, breaking down virtually all proteins into their individual amino acids; resolves carboxylic acids and alcohols.
Unit Definition
A unit of non-specific protease activity increases the rate of release of folin-positive amino acids and peptides from casein, at +40 °C and pH 7.5. For one unit (U) 1 μmol/min tyrosine is released, and for the unit "PU", the value is 1 μg/minute (1 U = 181 PU); for the unit "PUK", it is 0.1/minute (measured is the change in absorbance of molybdenum blue, formed by reaction with the folin ′s reagent under conditions such that 1 PUK = 50 PU).
Optimum temperature
35–40 °C
Stabilizers
Calcium ions: Calcium is highly effective in preventing pronase activity. The activity of a diluted solution containing 0.01–0.1 M calcium was stable over 24 hours at neutral pH at 4 to 8 °C. Pronase is also protected from heat inactivation by low levels of calcium.
Preparation Instructions
Working concentration: 0.5 to 2 mg/ml Working solution: Solvent is recommended in distilled water. Stock solution is prepared by adding pronase powder to distilled water (10 to 20 mg/ml). Storage conditions (working solution): -15 to -25 °C
Synonyms
non-specific protease
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