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P4850

Sigma-Aldrich

Proteinase K from Tritirachium album

buffered aqueous glycerol solution, for molecular biology, ≥800 units/mL

Synonym(s):

Endopeptidase K

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About This Item

CAS Number:
39450-01-6
Enzyme Commission number:
3.4.21.64 (BRENDA, IUBMB)
MDL number:
MFCD00132129
UNSPSC Code:
12352204
NACRES:
NA.54

grade

for 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

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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.
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.

Unit Definition

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).

Physical form

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

Legal Information

PicoGreen is a registered trademark of Life Technologies

pictograms

Health hazard
GHS08

signalword

Danger

hcodes

H334

Hazard Classifications

Resp. Sens. 1

Storage Class

10 - Combustible liquids

wgk_germany

WGK 2

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

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

Certificates of Analysis (COA)

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Robust Generation of Knock-in Cell Lines Using CRISPR-Cas9 and rAAV-assisted Repair Template Delivery
Vandemoortele G, et al.
Bio-protocol, 7(7), 1-6 (2017)
A S Neimanis et al.
Transboundary and emerging diseases, 65(1), 213-220 (2017-04-14)
Incursion of rabbit haemorrhagic disease virus (RHDV) into Sweden was documented in 1990 and it is now considered endemic in wild rabbit (Oryctolagus cuniculus) populations. Rabbit haemorrhagic disease virus 2 (RHDV2), a new, related lagovirus was first detected in France
Three-dimensional porous network structure developed in hydroxyapatite-based nanocomposites containing enzyme pretreated silk fibroin.
Wang Li, et al.
Journal of Nanoparticle Research, 6, 91-98 (2004)
Bovine herpes virus type 4 alters TNF-alpha and IL-8 profiles and impairs the survival of bovine endometrial epithelial cells
Chanrot M, et al.
Reproductive Biology, 17(3), 225-232 (2017)
K D Jany et al.
Biological chemistry Hoppe-Seyler, 366(5), 485-492 (1985-05-01)
The molecular mass of proteinase K was determined by gel electrophoresis in the presence of sodium dodecyl sulfate and by active site labelling with diisopropyl fluorophosphate. Both methods indicate molecular masses in the range of 27 000-29 000 Da. These
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Protocols

Proteinase K Enzymatic Assay

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

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