539480

Proteinase K from Tritirachium album

• 동의어(들): Endopeptidase K
• CAS Number: 39450-01-6
• 효소 위원회 번호: 3.4.21.14 (BRENDA, IUBMB)
• 효소 위원회 번호: 3.4.21.64 (BRENDA, IUBMB)
• MDL number: MFCD00132129
• UNSPSC 코드: 12352202
• NACRES: NA.21

속성

• 형태: lyophilized solid
• Quality Level: 400
• 특이 활성도: ≥30 mAnson units/mg dry wt; ≥40 mAnson units/mg protein
• 분자량: 28.93 kDa
• 제조업체/상표: Calbiochem^®
• 저장 조건: OK to freeze
• 색상: white
• solubility: 50 mM Tris-HCl, 2 mM calcium acetate, pH 8.0: soluble
• 외래 활성: 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)
• 배송 상태: ambient
• 저장 온도: 2-8°C

설명

일반 설명

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.

애플리케이션

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.

생화학적/생리학적 작용

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.

경고

Toxicity: Irritant (B)

단위 정의

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.

물리적 형태

Lyophilized from calcium acetate

재구성

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.

기타 정보

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.

법적 정보

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

안전성 정보

• 픽토그램: GHS08,GHS07
• 신호어: Danger
• 유해 및 위험 성명서: H315,H319,H334,H335
• 예방조치 성명서: P261 - P264 - P271 - P280 - P302 + P352 - P305 + P351 + P338
• Hazard Classifications: Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
• 표적 기관: Respiratory system
• Storage Class Code: 11 - Combustible Solids
• WGK: WGK 1
• Flash Point (°F): Not applicable
• Flash Point (°C): Not applicable