Proteinase K, RNase/DNase free

Product Description

Proteinase K is a highly reactive nonspecific serine protease that belongs to the subtilisin family of proteins. It cleaves at the carboxylic acid side of aliphatic, aromatic, or hydrophobic amino acids. Proteinase K is capable of inactivating RNases and DNases and is used in the isolation or preparation of high molecular weight nucleic acids. Proteinase K is also useful for helping to characterize enzymes, due to its cleavage specificity. This enzyme was designated proteinase K because of its ability to hydrolyze keratin. Proteinase K is stable in a wide variety of detergents and buffer salts and at various temperatures and pH. The isoelectric point of proteinase K is 8.9.

Proteinase K is typically used at a concentration of 50-100 μg/ml. It is active with or without the presence of SDS, urea, EDTA or various metal ions, but the activity of proteinase K can be increased by adding the denaturing agents and the structure of proteinase K can be stabilized by addition of Ca 2+. Proteinase K is inactivated by heating to 95°C for 10 minutes or using an inhibitor such as PMSF, AEBSF or DFP.

GoldBio's Proteinase K undergoes two sterile filtration steps during the separation and purification processes. The final product is further analyzed by plate culture to ensure the removal of all living organisms was 100% effective.

Product Specifications
Proteinase K, RNase/DNase free

MOLECULAR BIOLOGY GRADE

MW: 28.5 kDa

Activity: ≥30 U/mg

Unit Definition: One unit is defined as the amount of enzyme that will liberate 1.0 μmol tyrosine (Folin-positive amino acid) from casein per minute at 37°C, pH 7.5.

Source: Yeast cells with cloned gene encoding Engyodontium album (Tritirachium album) endolytic protease.

Storage/Handling: Store at -20°C.
Reconstitute in 50mM Tris-HCl (pH 8.0), 3mM CaCl2.


Proteinase K Quick Answers:

How do you inactivate proteinase K?

Inactivate proteinase K by heating it to 95 ˚C for 10 minutes. You can also permanently inactivate proteinase K by using protease inhibitors such as PMSF and AEBSF (Pefabloc®).


What is the best temperature for proteinase K activation?

The best temperature for proteinase K activity is between 50-65 ˚C. Higher temperatures help with protein unfolding, which makes it easier for proteinase K to breakdown proteins.


What is proteinase K used for?

Proteinase K is used in applications such as Digestion of unwanted proteins in molecular biology applications

  • Removal of endotoxins bound to cationic proteins such as lysozyme and RNaseA
  • Removal of nucleases for in situ hybridization
  • Prion research with respect to TSE (transmissible spongiform encephalopathies)
  • Protease footprinting
  • Mitochontrial isolation
  • Isolation of genomic DNA
  • Isolation of cytoplasmic RNA
  • Isolation of highly native DNA or RNA


How do you store proteinase K?

Aliquot proteinase K stock solution and store at -20 ˚C for up to 1 year.

Store lyophilized proteinase K powder desiccated at -20 ˚C for up to 2 years.


How do you dissolve proteinase K?

Proteinase K is water soluble and it can also be dissolved in Tris or PBS. Dissolving proteinase K with PBS is a little challenging, which may be due to pH. It's best to add proteinase K a little bit at a time while mixing into solution.


What is the optimal pH when working with proteinase K?

The optimal pH when working with proteinase K is between 7.5 - 12.0

Product Specifications

Catalog ID P-480
CAS # 39450-01-6
MW 28.5 kDa
Grade MOLECULAR BIOLOGY GRADE
Storage/Handling Store at -20°C.

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