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MilliporeSigma

539480

Sigma-Aldrich

Proteinase K from Tritirachium album

Synonym(s):

Endopeptidase K

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About This Item

CAS Number:
39450-01-6
EC Number:
3.4.21.14 (BRENDA, IUBMB)
EC Number:
3.4.21.64 (BRENDA, IUBMB)
MDL number:
MFCD00132129
UNSPSC Code:
12352202
NACRES:
NA.21

Key Documents

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form

lyophilized solid

Quality Level

400

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.
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.

Physical form

Lyophilized from calcium acetate

Preparation Note

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

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.
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.

Legal Information

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Toxicity: Irritant (B)

pictograms

Health hazardExclamation mark
GHS08,GHS07

signalword

Danger

Hazard Classifications

Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3

target_organs

Respiratory system

Storage Class

11 - Combustible Solids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Yujie Dong et al.
International journal of molecular sciences, 23(23) (2022-12-12)
Retinitis pigmentosa (RP) is a group of inherited retinal dystrophies that typically results in photoreceptor cell death and vision loss. Here, we explored the effect of early growth response-1 (EGR1) expression on photoreceptor cell death in Pde6brd1 (rd1) mice and
Richard P Tucker et al.
Methods in molecular biology (Clifton, N.J.), 2047, 421-437 (2019-09-26)
One of the first steps in studies of gene function is the spatiotemporal analysis of patterns of gene expression. Indirect immunohistochemistry is a method that allows the detection of a protein of interest by incubating a histological section with an
Markus Siegert et al.
Drug testing and analysis, 10(2), 243-253 (2017-09-08)
The vesicant sulfur mustard (SM) is a banned chemical warfare agent. Although, SM has been used in combat since WWI, there is no causal therapy currently available. Accordingly, development and investigation of antidotes and scavengers targeting SM are of high
Kongyang Zhu et al.
iScience, 25(12), 105636-105636 (2022-12-31)
A high point of Tibetan Plateau (TP) civilization, the expansive Tubo Empire (618-842 AD) wielded great influence across ancient western China. However, whether the Tubo expansion was cultural or demic remains unclear due to sparse ancient DNA sampling. Here, we
Jasmin Federizon et al.
Vaccine, 38(4), 942-950 (2019-11-16)
Outer surface protein A (OspA) is a Borrelia lipoprotein and an established Lyme disease vaccine target. Admixing non-lipidated, recombinant B. burgdorferi OspA with liposomes containing cobalt porphyrin-phospholipid (CoPoP) resulted in rapid, particulate surface display of the conformationally intact antigen. Particleization
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