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P2308

Sigma-Aldrich

Proteinase K from Tritirachium album

lyophilized powder, BioUltra, ≥30 units/mg protein, for molecular biology

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Synonym(s):
Endopeptidase K
CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
eCl@ss:
32160410
NACRES:
NA.54

grade

for molecular biology

Quality Level

product line

BioUltra

form

lyophilized powder

specific activity

≥30 units/mg protein

mol wt

28.93 kDa

composition

Protein, ≥90% biuret

impurities

≤1 ppm DNA

foreign activity

DNAse, Nickase and RNAse, none detected

shipped in

wet ice

storage temp.

−20°C

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This Item
P6556SRE0047SRE0005
form

lyophilized powder

form

lyophilized powder

form

lyophilized powder

form

buffered aqueous glycerol solution

specific activity

≥30 units/mg protein

specific activity

≥30 units/mg protein

specific activity

≥30 units/mg protein

specific activity

-

mol wt

28.93 kDa

mol wt

28.93 kDa

mol wt

28.93 kDa

mol wt

28.93 kDa

impurities

≤1 ppm DNA

impurities

-

impurities

≤1 ppm DNA

impurities

≤500 pg/mg DNA (PicoGreen® assay)

foreign activity

DNAse, Nickase and RNAse, none detected

foreign activity

Dnase ≤30 Kunitz units/mg solid, RNase ≤0.003 Kunitz units/mg solid

foreign activity

DNAse, Nickase and RNAse, none detected

foreign activity

DNAse, RNAse, exonuclease, endonuclease, and nickase, none detected

Application

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 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).

Pictograms

Health hazard

Signal Word

Danger

Hazard Statements

Hazard Classifications

Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2

Storage Class Code

11 - Combustible Solids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificates of Analysis (COA)

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MMYOMAG-74K-13

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Mamoru Oshiki et al.
Frontiers in microbiology, 9, 830-830 (2018-05-15)
Detection and genotyping of pathogenic RNA viruses in human and environmental samples are useful for monitoring the circulation and prevalence of these pathogens, whereas a conventional PCR assay followed by Sanger sequencing is time-consuming and laborious. The present study aimed
Margarita Zayas et al.
PLoS pathogens, 12(1), e1005376-e1005376 (2016-01-05)
Hepatitis C virus (HCV) nonstructural protein (NS)5A is a RNA-binding protein composed of a N-terminal membrane anchor, a structured domain I (DI) and two intrinsically disordered domains (DII and DIII) interacting with viral and cellular proteins. While DI and DII
Bahram Peivastegan et al.
BMC plant biology, 19(1), 262-262 (2019-06-19)
Stored potato (Solanum tuberosum L.) tubers are sensitive to wet conditions that can cause rotting in long-term storage. To study the effect of water on the tuber surface during storage, microarray analysis, RNA-Seq profiling, qRT-PCR and phytohormone measurements were performed
Mark F Sabbagh et al.
eLife, 7 (2018-09-07)
Vascular endothelial cell (EC) function depends on appropriate organ-specific molecular and cellular specializations. To explore genomic mechanisms that control this specialization, we have analyzed and compared the transcriptome, accessible chromatin, and DNA methylome landscapes from mouse brain, liver, lung, and
Sreenivas Adurthi et al.
Scientific reports, 7(1), 17289-17289 (2017-12-13)
Oestrogen controls Foxp3 expression in regulatory T cells (Treg cells) via a mechanism thought to involve oestrogen receptor alpha (ERα), but the molecular basis and functional impact of ERα signalling in Treg cells remain unclear. We report that ERα ligand

Articles

An Introduction to Proteinase K and Its Applications

Proteinase K is commonly used in molecular biology and biochemistry applications to digest structural proteins and enzymes. It is useful in removing nucleases that can degrade DNA and RNA, as well as in the isolation of intact genomic DNA from various sources.

How Proteinase K Can Be Used in Tissue Samples

The use of Pro K in combination with other reagents, such as detergents and chaotropic agents, can help to disrupt the cell membranes and release DNA from tissue. This is particularly important for downstream applications such as PCR, sequencing, and other molecular biology techniques that require pure and intact DNA.

How to Determine the Best Product & Supplier for Proteinase K

The cost of proteinase K can vary depending on the source, purity, manufacturing process and vendor’s quality management system. It is important to balance the cost with the desired quality, performance, documentation and technical/quality support to select the optimal Proteinase K for the intended application

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.

See All

Protocols

GenElute™ Blood Genomic DNA Kit Protocol

The GenElute™ Blood Genomic DNA Kit Protocol provides a simple and convenient way to isolate pure genomic DNA from fresh or aged whole blood.

GenElute™ Mammalian Genomic DNA Miniprep Kit Protocol

The GenElute Mammalian Genomic DNA Purification Kit Protocol describes the isolation of pure, high molecular weight DNA from a variety of mammalian sources.

Enzymatic Assay of Proteinase K (EC 3.4.21.64)

Proteinase K (EC 3.4.21.64) activity can be measured spectrophotometrically using hemoglobin as the substrate. Proteinase K hydrolyzes hemoglobin denatured with urea, and liberates Folin-postive amino acids and peptides. One unit will hydrolyze hemoglobin to produce color equivalent to 1.0 μmol of tyrosine per minute at pH 7.5 at 37 °C (color by Folin & Ciocalteu's Phenol Reagent).

GenElute™ Bacterial Genomic DNA Kit Protocol (NA2100, NA2110, NA2120)

GenElute™ Bacterial Genomic DNA Kit protocol describes a simple and convenient way for the isolation of pure genomic DNA from bacteria.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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