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PRON-RO

Roche

Pronase

from Streptomyces griseus

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Synonym(s):
non-specific protease
Enzyme Commission number:
3.4.24.4 (BRENDA, IUBMB)

biological source

Streptomyces griseus

Quality Level

100

sterility

non-sterile

form

lyophilized powder

specific 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.)

packaging

pkg of 1 g (10165921001)
pkg of 5 g (11459643001)

manufacturer/tradename

Roche

concentration

0.5-2 mg/mL

parameter

35-40 °C optimum reaction temp.

technique(s)

DNA extraction: suitable

color

white

optimum pH

7.0-8.0

pH range

6.0-9.0

solubility

water: soluble 10-20 g/L

suitability

suitable for molecular biology

NCBI accession no.

WP_012381357.1

UniProt accession no.

B1W0S5
P00776

application(s)

life science and biopharma

foreign activity

Deoxyribonuclease <0.002 U/mg ( based on lyoph.)
Ribonuclease <0.002 U/mg ( based on lyoph.)

storage temp.

2-8°C

Gene Information

Streptomyces griseus ... SGR_RS27460(6214424)

Related Categories

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).

Storage and Stability

Store dry

Other Notes

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

Pictograms

Exclamation markHealth hazard
GHS07,GHS08

Signal Word

Danger

Hazard Statements

H315 - H319 - H334 - H335

Hazard Classifications

Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3

Target Organs

Respiratory system

Storage Class Code

11 - Combustible Solids

WGK

WGK 1

Flash Point(F)

does not flash

Flash Point(C)

does not flash

Regulatory Information

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