Proteinase K Solution 20mg/ml
|Molecular Biology||Storage: -20°CShipping: Blue Ice|
Proteinase K is classified as a non-specific serine protease. It is an endolytic protease that cleaves peptide bonds at the carboxylic sides of aliphatic, aromatic or hydrophobic amino acids.
• Isolation of high molecular weight DNA
• Isolation of plasmid and genomic DNA
• Isolation of RNA
• Inactivation of RNase and DNase activities
• Ready to use, provided in liquid form containing 40% glycerol.
• Smaller contamination risk with liquid form.
• Can be stored at -20°C still having the enzyme in liquid form.
• Excellent stability
•Proteinase K is not inactivated by metal-chelating agents like EDTA, by thiol-reactive reagents (such as monoiodoacetic acid, p-chloromercuribenzoate) or by specific trypsin and chymotrypsin inhibitors. Phenylmethylsulfonyl fluoride and diisopropyl phosphorofluoridate completely inhibit the enzyme.
•Proteinase K is stable over a wide pH range: 4-12.5.
•Raising the temperature of the reaction from 37°C to 50°-65°C can increase the activity of Proteinase K.
•Proteinase K can be inactivated at 95°C for 10 minutes (some enzymatic activity may still remain)
•Proteinase K retains its activity in the presence of SDS (0.5-1%) or urea (1-4M).
•Depending on the application the enzyme is used in the absence or presence of 0.2-1% SDS or 1-4M urea.
Nomenclature: EC 22.214.171.124
Recommended Dilution: The recommended working concentration for Proteinase K is 0.05-1mg/ml. Use 3ul of a 20mg/ml solution per 1.5ml of bacterial culture.
Optimal dilutions to be determined by researcher.
Clear, colorless, complete
pH: 7.5 ± 0.2
One unit liberates 1umole of Folin positive amino acids, measured as Tyrosine, at 37°C, pH 7.5, using urea-denatured hemoglobin as the substrate.
Nicking Activity: None Detected
RNases: None Detected
Exonucleases: None Detected
Phosphatases: None Detected
Storage and Stability:
May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Source: Tritirachium album limber
Form: Supplied as a liquid in 10mM Tris HCl, pH 7.5, 1mM calcium acetate, 40% glycerol.
Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
US Biological application reference: Neale, J.R. et al., (2009) Polycycl Aromat Compd. 28:402–417. 1. Ebeling, W., et al., Eur. J. Biochem 47: 91-97 (1974). 2. Ausubel, F.M., et al., Current Protocols in Molecular Biology,, John Wiley (1992). 3. Maniatis, T., et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press (1989). 4. Wiegers, U., Hilz, H., Biochem. and Biophys. Res. Commun. 44: 513-519 (1971). 5. Hilz, H., et al., Eur. J. Biochem. 56: 103-108 (1975). 6. Brdiczka, D., Krebs, W., Biochim. Biophys. Acta 297: 203-212 (1973). 7. Crowe, J.S., et al., Nucleic Acids Res. 19: 184, (1991).