Proteinase K Solution 20mg/ml

Blue Ice

28, 9

Supplied as a liquid in 10mM Tris HCl, pH 7.5, 1mM calcium acetate, 40% glycerol.

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.

Applications:
• Isolation of high molecular weight DNA
• Isolation of plasmid and genomic DNA
• Isolation of RNA
• Inactivation of RNase and DNase activities

Advantages:
• 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

Notes:
•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=3.4.21.64; PROK; Endopeptidase K; Tritirachium Alkaline Proteinase

Source:
Tritirachium album limber

Concentration:
~20mg/ml

Form:
Supplied as a liquid in 10mM Tris HCl, pH 7.5, 1mM calcium acetate, 40% glycerol.

Recommended Dilutions:
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.

Solubility:
Colorless to pale yellow, clear, complete

pH:
7.5 ± 0.2

Specific Activity:
≥30 units/mg

Unit Definition:
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.

Quality Control:
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 6 months after receipt 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.