SAE0151

Proteinase K aus Tritirachium album

free of DNA contaminants, suitable for Microbiome research, lyophilized powder, ≥30 units/mg protein

• Synonym(e): Proteinase K aus Tritirachium album, Endopeptidase K
• CAS-Nummer: 39450-01-6
• EC-Nummer: 3.4.21.64 (BRENDA, IUBMB)
• MDL-Nummer: MFCD00132129
• UNSPSC-Code: 12352202
• NACRES: NA.77

Eigenschaften

• Qualitätsniveau: 200
• Spezifische Aktivität: 30 units/mg protein
• Mol-Gew.: 28.93 kDa
• Leistungsmerkmale: DNA free
• Versandbedingung: wet ice
• Lagertemp.: −20°C

Beschreibung

Allgemeine Beschreibung

Purified Proteinase K DNA free SAE0151 undergoes strict quality control testing to ensure the absence of detectable levels of contaminating DNA using 35 cycles PCR amplification of 16S and 18S rDNA using universal primer sets.

Proteinase K is a stable serine protease with broad substrate specificity. It degrades many proteins in the native state even in the presence of detergents. Proteinase K was isolated from a fungus able to grow on keratin and the enzyme can digest native keratin. The enzyme belongs to the subtilisin family with an active site catalytic triad (Asp39 -His69 -Ser224).
The predominant site of cleavage is the peptide bond adjacent to the carboxyl group of aliphatic and aromatic amino acids with blocked alpha amino groups. It is commonly used for its broad specificity. Proteinase K is usually denatured by subsequent phenol extractions.

The study of microbial communities has been revolutionized in recent years by the widespread adoption of culture independent analytical techniques such as 16S rRNA gene sequencing and metagenomics. Since DNA contamination during sample preparation is a major problem of these sequence-based approaches, DNA extraction reagents free of DNA contaminates are essential.

Anwendung

Nützlich zur proteolytischen Inaktivierung von Nukleasen bei der Isolierung von DNA und RNA.
Entfernt Endotoxine, die an kationische Proteine wie Lysozym und Ribonuklease A binden.
Berichten zufolge nützlich für die Isolierung von Leber-, Hefe- und Mungbohnen-Mitochondrien
Bestimmung der Enzymlokalisierung auf Membranen
Behandlung von Paraffinschnitten zur Freilegung von Antigenbindungsstellen zur Antikörpermarkierung.
Verdauung von Proteinen aus Hirngewebeproben für Prionen in der TSE-Forschung.

• Proteinase K is frequently used in molecular biology applications to digest unwanted proteins, such as nucleases from DNA or RNA preparations from microorganisms, cultured cells, and plants.
• Removes endotoxins that bind to cationic proteins such as lysozyme and ribonuclease A 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. Wirkung

Proteinase K ist eine stabile und hochreaktive Serinprotease. Ergebnisse von Kristall- und Molekularstrukturstudien lassen darauf schließen, dass das Enzym zur Subtilisin-Familie mit einer katalytischen Triade (Asp³⁹-His⁶⁹-Ser²²⁴) im aktiven Zentrum gehört. Es ist in einer Reihe von Umgebungen stabil: pH, Puffersalze, Detergenzien (SDS) und Temperatur. In Gegenwart von 0,1–0,5 % SDS bleibt Proteinase K aktiv und verdaut zahlreiche Proteine und Nukleasen in DNA-Präparationen, ohne die Integrität der isolierten DNA zu beeinträchtigen.

Leistungsmerkmale und Vorteile

Purified Proteinase K DNA free SAE0151 undergoes strict quality control testing to ensure it will be free of DNA contaminants, suitable for Microbiome research.

Einheitendefinition

One unit will hydrolyze urea denatured hemoglobin to produce color equivalent to 1.0 micromole of tyrosine per minute at pH 7.5, 37°C (color per Folin-Ciocalteu reagent).

Angaben zur Herstellung

Proteinase K is active in 1% Triton X-100 and in 0.5% (w/v) SDS. SDS and urea will denature protein substrates resulting in increased digestion rates. Proteinase K itself is denatured much more slowly by these agents.

Sicherheitshinweise

• Piktogramme: GHS08,GHS07
• Signalwort: Danger
• H-Sätze: H315,H319,H334,H335
• P-Sätze: P261 - P264 - P271 - P280 - P302 + P352 - P305 + P351 + P338
• Gefahreneinstufungen: Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
• Zielorgane: Respiratory system
• Lagerklassenschlüssel: 11 - Combustible Solids
• WGK: WGK 1
• Flammpunkt (°F): Not applicable
• Flammpunkt (°C): Not applicable