539480
Proteinase K from Tritirachium album
Synonym(s):
Endopeptidase K
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About This Item
CAS Number:
MDL number:
UNSPSC Code:
12352202
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form
lyophilized solid
Quality Level
specific activity
≥30 mAnson units/mg dry wt
≥40 mAnson units/mg protein
mol wt
28.93 kDa
manufacturer/tradename
Calbiochem®
storage condition
OK to freeze
color
white
solubility
50 mM Tris-HCl, 2 mM calcium acetate, pH 8.0: soluble
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
ambient
storage temp.
2-8°C
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General description
Serine protease that exhibits strong proteolytic activity on a wide variety of denatured and native proteins of high molecular weight. Because of its rapid proteolytic inactivation of endogenous nucleases, it can be used for isolation of mRNA and high molecular weight DNA. Preferentially cleaves bonds next to the carbonyl group of N-substituted hydrophobic, aliphatic, and aromatic amino acids. Inhibited by DFP, Hg2+, and PMSF. Not inactivated by metal chelators, sulfhydryl reagents, TLCK, or TPCK.
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.
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.
Warning
Toxicity: Irritant (B)
Unit Definition
One mAnson unit is defined as the amount of enzyme that will liberate 1.0 µmol of Folin-positive amino acids (calculated as tyrosine) per min at 37°C, pH 7.5.
Physical form
Lyophilized from calcium acetate
Reconstitution
Following reconstitution, store in the refrigerator (4°C). Stock solutions prepared in 50 mM Tris-HCl, 2 mM calcium acetate, pH 8.0 are stable for up to 1 year at 4°C.
Other Notes
Watazu, Y., et al. 1993. J. Lab. Clin. Anal. 7, 81.
Lebherz, H.G., et al. 1986. Biochem. J.233, 51.
Ebeling, W., et al. 1974. Eur. J. Biochem.47, 91.
Lebherz, H.G., et al. 1986. Biochem. J.233, 51.
Ebeling, W., et al. 1974. Eur. J. Biochem.47, 91.
Legal Information
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
Signal Word
Danger
Hazard Statements
Precautionary Statements
Hazard Classifications
Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2
Storage Class Code
11 - Combustible Solids
WGK
WGK 1
Certificates of Analysis (COA)
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