SAE0151

Proteinasa K from Tritirachium album

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

• Sinónimos: Endopeptidasa K
• Número de CAS: 39450-01-6
• Comisión internacional de enzimas: 3.4.21.64 (BRENDA, IUBMB)
• MDL number: MFCD00132129
• UNSPSC Code: 12352202
• NACRES: NA.77

Propiedades

• Quality Level: 200
• specific activity: 30 units/mg protein
• mol wt: 28.93 kDa
• feature: DNA free
• shipped in: wet ice
• storage temp.: −20°C

Descripción

General description

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.

Application

Útil para la inactivación proteolítica de nucleasas durante el aislamiento de ADN y ARN.
Elimina las endotoxinas que se unen a las proteínas catiónicas como la lisozima y la ribonucleasa A.
Se ha comunicado que es útil para el aislamiento de mitocondrias hepáticas, de levadura y de frijol mung
Determinación de la localización de las enzimas en las membranas
Tratamiento de cortes de tejido incluidos en parafina para exponer los sitios de unión de antígenos para el marcado con anticuerpos.
Digestión de proteínas de muestras de tejido cerebral para investigación de los priones en las encefalopatías espongiformes transmisibles (EET).

• 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 Actions

La proteinasa K es una serín proteasa estable y muy reactiva. La evidencia procedente de estudios de la estructura cristalina y molecular indica que la enzima pertenece a la familia subtilisina con una tríada catalítica en el sitio activo (ASP³⁹-HIS⁶⁹-ser²²⁴). Es estable en una amplia variedad de entornos: pH, sales amortiguadoras, detergentes (SDS) y temperatura. En presencia de SDS al 0,1 - 0,5 %, la proteinasa K conserva su actividad y digerirá una variedad de proteínas y nucleasas en preparaciones de ADN sin comprometer la integridad del ADN aislado.

Features and Benefits

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

Unit Definition

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).

Preparation Note

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.

Información de seguridad

• pictograms: GHS08,GHS07
• signalword: Danger
• hcodes: H315,H319,H334,H335
• pcodes: P261 - P264 - P271 - P280 - P302 + P352 - P305 + P351 + P338
• 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