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  • P6556
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P6556

Sigma-Aldrich

Proteinase K from Tritirachium album

lyophilized powder, ≥30 units/mg protein

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

form

lyophilized powder

Quality Level

specific activity

≥30 units/mg protein

mol wt

28.93 kDa

solubility

H2O: soluble 1 mg/mL, clear, colorless

foreign activity

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

shipped in

wet ice

storage temp.

−20°C

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This Item
SAE0009P5568539480
form

lyophilized powder

form

lyophilized powder

form

buffered aqueous glycerol solution

form

lyophilized solid

specific activity

≥30 units/mg protein

specific activity

≥30 units/mg protein

specific activity

-

specific activity

≥30 mAnson units/mg dry wt, ≥40 mAnson units/mg protein

mol wt

28.93 kDa

mol wt

28.93 kDa

mol wt

28.93 kDa

mol wt

28.93 kDa

solubility

H2O: soluble 1 mg/mL, clear, colorless

solubility

-

solubility

-

solubility

50 mM Tris-HCl, 2 mM calcium acetate, pH 8.0: soluble

foreign activity

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

foreign activity

DNAse, RNAse, none detected.

foreign activity

-

foreign activity

DNase, none detected (nicking activity with pBR322, incubation for 6 h at 37°C), RNase, none detected (ribonuclease activity with MS2 RNA detected after incubation for 6 h at 37°C)

shipped in

wet ice

shipped in

-

shipped in

-

shipped in

ambient

Application

Product P6556 is provided as a lyophilized powder. Product P6556 has been used to break down human lens protein. Protease footprinting by Proteinase K digestion can reveal protein-protein surface interactions. The enzyme from Sigma has been used in the pre-hybridization step of chicken embryos. It has also been used for the enrichment of PrPSc, a prion protein that is present in sheep, hamster and mouse scrapie samples.
Proteinase K is useful for the proteolytic inactivation of nucleases during the isolation of DNA and RNA.
It is used for the removal of endotoxins bound to cationic proteins such as lysozyme and ribonuclease A.
It is useful for the isolation of hepatic, yeast, and mung bean mitochondria
and is used to determine enzyme localization on membranes
It is used for the treatment of paraffin embedded tissue sections to expose antigen binding sites for antibody labeling and
for digestion of proteins from brain tissue samples for prions in Transmissible Spongiform Encephalopathies (TSE) research. Product P6556 is provided as a lyophilized powder. Product P6556 has been used to break down human lens protein.
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 optimum pH is between 7.5-9.0 and the isoelectric point is 8.9 Ca2+ (1-5 mM) is required for activation. Proteinase K is inhibited by diisopropyl fluorophosphate (DFIP), and phenylmethanesulfonyl fluoride (PMSF).
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

Certificate of Analysis

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Certificate of Origin

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