Proteinase K – 200mg
- Used for digestion of the protein keratin
Proteinase K (Pro K) is a stable >50 mg/ml (powder) highly active proteolytic enzyme that is purified from the mold Tritirachium album. The enzyme has two binding sites for Ca++, which lie some distance from the active site and are not directly involved in the catalytic mechanism. The high activity of the enzyme solution (0.6 U/µl) means that you can keep the reaction volumes low (as low as 200 µg/ml). It also means that you can expect more complete digestion of your samples. Used primarily to digest protein and remove contamination from nucleic acid sample preps. The addition of pro-k to nucleic acid preparations rapidly inactivates nucleases that might otherwise degrade the DNA or RNA during purification.
Click on Technical Information tab for more detailed info and solution concentration recipe.
Molecular Weight: ~18,000 Da
Activity @37°C (U/mg) Min.: 30 U/mg
Molecular Biology Specifications
DNase Assay: None Detected
RNase Assay: None Detected
Proteinase K is a stable >50 mg/ml (power) highly active proteolytic enzyme that is purified from the mold tritirachium album. The enzyme has two binding sites for Ca++, which lie some distance from the active site and is not directly involved in the catalytic mechanism. The high activity of the enzyme solution (0.6 U/μl) means that you can keep the reaction volumes low, for instance, as low as 200 μg/ml. It also means that you can expect more complete digestion of your sample.
To Make a 20 mg/ml Proteinase K Solution (Conc.) TRIS: 10 mM Calcium Chloride: 1 mM Glycerol: 30%
pH adjusted to 8.0 with HCl
Storage Store at -20°C. Warm to room temperature before opening. Keep tightly sealed.
How do I deactivate the Proteinase K in my sample? Heat the sample to 95°C for 20-30 minutes.
Why did the Proteinase K solution get cloudy after being stored for a week? Why did the Proteinase K solution form precipitates after being stored for a week? A number of factors can be responsible for these conditions. 1)If EDTA is used in the buffer solution it can cause chelating of the calcium and magnesium causing cloudiness 2)Customer may need to mix fresh solution each time they use Pro-K in their reactions. 3)If no EDTA is used and precipitates form after storing, activity is not affected, re-dissolve precipitates. 4)Recommended solution is 20 mg/ml, if mixed at a greater concentration, material can go out of solution. Put back in solution, activity not affected.
How is the Proteinase K solution purified? The enzyme is purified by chromatography.
What is the best buffer solution to use for Proteinase K use? Calcium is a key ingredient for maintaining viability of the enzyme once in solution. The ideal buffer solution is as follows: 50 mM TRIS, pH 8.3 3 mM CaCl2 50% glycerol Solution can be stored at -20°C with minimal freeze/thaw cycles ideally. Otherwise 2° – 8°C is also acceptable.