P6556

Proteinasa K from Tritirachium album

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)
• EC Number: 254-457-8
• MDL number: MFCD00132129
• UNSPSC Code: 12352204
• eCl@ss: 32160410
• NACRES: NA.54

Propiedades

• form: lyophilized powder
• Quality Level: 300
• specific activity: ≥30 units/mg protein
• mol wt: 28.93 kDa
• technique(s): DNA extraction: suitable
• solubility: H₂O: soluble 1 mg/mL, clear, colorless
• foreign activity: Dnase ≤30 Kunitz units/mg solid; RNase ≤0.003 Kunitz units/mg solid
• shipped in: wet ice
• storage temp.: −20°C

Descripción

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).

Product P6556 is provided as a lyophilized powder. Product P6556 has been used to break down human lens protein. Protease footprinting by Proteinase K digestion can reveal protein-protein surface interactions. The enzyme from Sigma has been used in the pre-hybridization step of chicken embryos. It has also been used for the enrichment of PrP^Sc, a prion protein that is present in sheep, hamster and mouse scrapie samples.

Proteinase K is useful for the proteolytic inactivation of nucleases during the isolation of DNA and RNA.
It is used for the removal of endotoxins bound to cationic proteins such as lysozyme and ribonuclease A.
It is useful for the isolation of hepatic, yeast, and mung bean mitochondria
and is used to determine enzyme localization on membranes
It is used for the treatment of paraffin embedded tissue sections to expose antigen binding sites for antibody labeling and
for digestion of proteins from brain tissue samples for prions in Transmissible Spongiform Encephalopathies (TSE) research. Product P6556 is provided as a lyophilized powder. Product P6556 has been used to break down human lens protein.

Biochem/physiol Actions

Proteinase K has a broad specificity and degrades many proteins even in the native state. It mainly cleaves the peptide bond adjacent to the carboxyl group of aliphatic and aromatic amino acids with blocked α-amino groups. The optimum pH is between 7.5-9.0 and the isoelectric point is 8.9 Ca²⁺ (1-5 mM) is required for activation. Proteinase K is inhibited by diisopropyl fluorophosphate (DFIP), and phenylmethanesulfonyl fluoride (PMSF).

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.

Unit Definition

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

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
• ppe: dust mask type N95 (US), Eyeshields, Faceshields, Gloves