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P4850

Proteinase K from Tritirachium album

Name: Proteinase K from <I>Tritirachium album</I>
Brand: SIGMA

buffered aqueous glycerol solution, Molecular Biology, ≥800 units/mL

Synonym(s):

Endopeptidase K

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About This Item

CAS Number:

39450-01-6

UNSPSC Code:

12352204

NACRES:

NA.54

EC Number:

3.4.21.64 (BRENDA, IUBMB)

MDL number:

MFCD00132129

eCl@ss:

32160410

Concentration:

≥10 mg/mL, ≥800 units/mL

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Properties

grade

Molecular Biology

Quality Level

200

form

buffered aqueous glycerol solution

mol wt

28.93 kDa

concentration

≥10 mg/mL, ≥800 units/mL

impurities

≤0.5 ppm DNA (PicoGreen^® assay)

foreign activity

DNase, Nickase and RNase, none detected

storage temp.

2-8°C

Description

Application

Proteinase K from Tritirachium album has been used:

in bovine endometrial epithelial cells for herpes viral DNA extraction
for viral RNA extraction from nasal swabs
as a component of phase lock and direct PCR lysis buffer

The product has been used to study its pre-treatment effects on the silk fibroin. The aspects analysed in this study included the crystallographic properties of hydroxyapatite (HAp), and the microstructure and microhardness of the composites. The enzyme has also been used to facilitate the access of probes to rRNA using FISH techniques to detect pathogenic Staphylococcus aureus.

Useful for the proteolytic inactivation of nucleases during the isolation of DNA and RNA.

Useful for the proteolytic inactivation of nucleases during the isolation of DNA and RNA.
Removes endotoxins that bind to cationic proteins such as lysozyme and ribonuclease A.
Reported useful for the isolation of hepatic, yeast, and mung bean mitochondria
Determination of enzyme localization on membranes
Treatment of paraffin embedded tissue sections to expose antigen binding sites for antibody labeling.
Digestion of proteins from brain tissue samples for prions in Transmissible Spongiform Encephalopathies (TSE) research.

Biochem/physiol Actions

Proteinase K has a broad specificity and degrades many proteins even in the native state. It mainly cleaves the peptide bond adjacent to the carboxyl group of aliphatic and aromatic amino acids with blocked α-amino groups. The molecular weight of proteinase K from amino acid sequence is found to be 28,930 Da and from SDS-PAGE, it is found to be 28,500 Da. The optimum pH is between 7.5-9.0 and its isoelectric point is 8.9. Ca²⁺ (1-5 mM) is required for its activation. Proteinase K is inhibited by DIFP (diisopropylfluorophosphate) or PMSF (phenylmethylsulfonyl fluoride).

Proteinase K is a stable and highly reactive serine protease. Evidence from crystal and molecular structure studies indicates the enzyme belongs to the subtilisin family with an active-site catalytic triad (Asp³⁹-His⁶⁹-Ser²²⁴). It is stable in a broad range of environments: pH, buffer salts, detergents (SDS), and temperature. In the presence of 0.1-0.5% SDS, proteinase K retains activity and will digest a variety of proteins and nucleases in DNA preparations without compromising the integrity of the isolated DNA.

Physical form

Solution in 40% glycerol (v/v) containing 10 mM Tris-HCl, pH 7.5, with 1 mM calcium acetate.

Other Notes

One unit will hydrolyze urea-denatured hemoglobin to produce color equivalent to 1.0 μmole of tyrosine per min at pH 7.5 at 37 °C (color by Folin-Ciocalteu reagent).

Legal Information

PicoGreen is a registered trademark of Life Technologies

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pictograms

GHS08

signalword

Danger

hcodes

H334

pcodes

P261 - P284 - P501

Hazard Classifications

Resp. Sens. 1

Storage Class

10 - Combustible liquids

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

Eyeshields, Faceshields, Gloves, type ABEK (EN14387) respirator filter

Regulatory Information

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Protocols

Proteinase K Enzymatic Assay

Proteinase K activity measured via spectrophotometry using hemoglobin substrate, crucial for enzyme characterization.

Articles

Best Product & Supplier for Proteinase K

Balancing Proteinase K cost with quality and technical support ensures optimal enzyme selection for diverse applications.

How Proteinase K Can Be Used in Blood DNA Extraction Applications

In blood DNA extraction, Proteinase K, an enzyme commonly used to degrade proteins, can help break down the cellular and nuclear membranes, releasing DNA from the cells that protect it from degradation and increase purity/yield making it more suitable for various molecular biology techniques.

Guide on How to Use Proteinase K in Different Procedures

Guidelines on use of proteinase K, an enzyme commonly used to degrade proteins, and protect DNA and RNA from degradation in samples.

View All Articles

Global Trade Item Number

SKU	GTIN
P4850-.025ML	04061824821079
P4850-.110ML	04061824821093
P4850-1ML	04061824821147
P4850-300UL	04061824821161
P4850-5ML	04061824821192
P4850-100UL	04061824821116
P4850-10UL	04061824821123
P4850-50UL	04061824821178