Proteinase K, RNase/DNase free
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.
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
|Grade||MOLECULAR BIOLOGY GRADE|
|Storage/Handling||Store at -20°C.|
A general protocol for inactivating nucleases with Proteinase K during the extraction of nucleic acids
A protocol for the extraction of DNA from mouse tail using Proteinase K
Your free, printable guide to proteinase K, with general info, usage details and how to activate and optimize.
GoldBio Proteinase K efficacy and analyses; including DNase/RNase contamination assays and DNA extraction assays.
The Proteinase K Digestion Guide includes 4 tables showing the experimental setup for proteinase K digestion and includes the referenced publication.
This handbook is packed with information about proteinase K, its applications, protocols, troubleshooting, FAQs and more. If you're working with proteinase K, you don't want to not have this handbook.
Sample Certificate of Analysis - Proteinase K
COA for example only. Actual tests may differ.