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Key Documents

70663

Millipore

Proteinasa K from Tritirachium album

Highly active serine protease that exhibits broad cleavage specificity on native and denatured proteins and is widely used in the purification of DNA and RNA.

Sinónimos:

Endopeptidasa K

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About This Item

Número de CAS:
Comisión internacional de enzimas:
MDL number:
UNSPSC Code:
12352202
NACRES:
NA.85

Quality Level

form

solid

mol wt

28.93 kDa

manufacturer/tradename

Novagen®

storage condition

OK to freeze

technique(s)

DNA extraction: suitable

shipped in

wet ice

storage temp.

2-8°C

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General description

Proteinase K is a highly active 28,904-Daserine protease isolated from the fungusTritirachium album. The enzyme exhibitsbroad cleavage specificity on native anddenatured proteins and is widely used in thepurification of DNA and RNA. Its activityis increased in the presence of denaturantssuch as SDS (1%) and elevated temperature(50-60°C). The recommended workingconcentration is 50-100 µg/ml for proteinremoval and enzyme inactivation, and up to2 mg/ml for tissue treatment. The Proteinase K,Lyophilized powder can be prepared as a20 mg/ml stock solution in water and storedin aliquots at -20°C. The enzyme is alsoavailable as a ready-to-use concentrated stocksolution (600 mAU/ml) that is convenientfor routine use in most applications. 1 mgof Proteinase K is the equivalent of 30 mAU(AU = Anson unit). The Novagen ProteinaseK products are free of detectable DNase andRNase

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).

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.

Warning

Toxicity: Harmful (C)

Unit Definition

One AU (AU = Anson unit) is defined as the amount of enzyme that liberates 1.0 µmol (181 µg) of tyrosine from casein per minute at pH 7.5 at 37°C.

Legal Information

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

pictograms

Health hazardExclamation mark

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


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