Is proteinase K Stable at room temperature?

QIAGEN Proteinase K is stable for up to 1 year after delivery when stored at room temperature. To prolong the shelf-life of Proteinase K, storage at 2–8°C is recommended.

What temperature is proteinase K activity?

between 20 and 60°C
Proteinase K is active in a wide range of temperatures and buffers with optimal activity between 20 and 60°C and a pH between 7.5 and 12.0 (1, 2). Activity is stimulated when up to 2% SDS or up to 4 M urea are included in the reaction (3).

Why do we inactivate proteinase K?

For example, in the nucleic acid extraction protocol, proteinase K is added to cell lysate and then an incubation period follows to ensure a complete digestion. To prevent potential digestion of your samples, proteinase K is inactivated after incubation. The common temperature for inactivation is 95°C.

What’s the best way to inactivate proteinase K?

The most effective way to inactivate the enzyme, as with most proteins is to increase the temperature or change the pH significantly. Proteinase K is inactivated by heat (e.g. incubating at 55°C). How do you determine if the enzyme is working?

How does temperature affect the activity of Proteinase K?

The above-mentioned conditions enhance proteinase K activity by making its substrate cleavage sites more accessible. Temperatures above 65 °C, trichloroacetic acid (TCA) or the serine protease-inhibitors AEBSF, PMSF or DFP inhibit the activity.

What is the shelf life of Proteinase K?

It has a shelf life of 12 months when stored in a dry place at 4–8°C, due to the fact that it is a very stable. Short term storage at ambient temperatures do not harm the enzyme’s activity and stability.

How to isolate RNA with Proteinase K and SDS?

Isolation of RNA: For cytoplasmic RNA isolation, centrifuge the cell lysate, remove the supernate and add 200 ug/ml proteinase K and SDS to 2% (w/v). Incubate for 30 minutes at 37°C.