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P2308

Sigma-Aldrich

Proteinase K from Tritirachium album

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

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)
EC Number:
254-457-8
MDL number:
MFCD00132129
UNSPSC Code:
12352204
eCl@ss:
32160410
NACRES:
NA.54

grade

for molecular biology

Quality Level

400

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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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).

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Pictograms

Health hazardExclamation mark
GHS08,GHS07

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)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

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Certificates of Analysis (COA)

Lot/Batch Number
SLBX1525
089M4033V
50180408
SLCH8204
SLCH8209

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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
Ruobing Jia et al.
Molecular cancer, 18(1), 161-161 (2019-11-15)
Dynamic N6-methyladenosine (m6A) RNA modification generated and erased by N6-methyltransferases and demethylases regulates gene expression, alternative splicing and cell fate. Ocular melanoma, comprising uveal melanoma (UM) and conjunctival melanoma (CM), is the most common primary eye tumor in adults and
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
Jimmy Su et al.
Journal of biomedical materials research. Part A, 106(9), 2448-2462 (2018-04-18)
Hydrogels, highly-hydrated crosslinked polymer networks, closely mimic the microenvironment of native extracellular matrix (ECM) and thus present as ideal platforms for three-dimensional cell culture. Hydrogels derived from tissue- and organ-specific decellularized ECM (dECM) may retain bioactive signaling cues from the
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Articles

Enzymatic Assay of Proteinase K with Hemoglobin Substrate

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).

Protocols

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.

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.

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.

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