PRONASE FROM STREPTOMYCES GRISEUS

General description

Pronase is a group of proteolytic enzymes derived from Streptomyces griseus K-1. It consists of various components, including serine-type proteases, Zn2+ endopeptidases, Zn²⁺-leucine aminopeptidases, and Zn²⁺ carboxypeptidases.

Pronase is a mixture of several nonspecific endo- and exoproteases that digest proteins down to single amino acids.

Specificity

Pronase has a broad specificity, breaking down virtually all proteins into their individual amino acids; resolves carboxylic acids and alcohols.

Application

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. It has been used for the dechorionation of Zebrafish embryos.

Biochem/physiol Actions

Pronase has wide-ranging specificity. Therefore, it is commonly used when extensive or complete degradation of proteins is required. Its applications include protein analysis in cell organelles, removal of proteins during DNA and RNA isolation, and the production of protein hydrolysates for amino acid analysis. In addition, Pronase surpasses trypsin in its ability to disperse fibroblastic cell lines, excelling in both monolayer detachment and the creation of single-cell suspensions.

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).

Preparation Note

Stabilizers: Protective effect of calcium ions

The preparation contains 20% calcium acetate for stability. It is free of starch as per the current Quality Control Procedures.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.
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

Reconstitution

The lyophilizate is soluble in water (10 to 20 mg/ml).

Other Notes

For life science research only. Not for use in diagnostic procedures.