Feb 10, 2022
Version 2
Thermolabile Proteinase K Typical Reaction Protocol V.2
- New England Biolabs1
- 1New England Biolabs
- New England Biolabs (NEB)Tech. support phone: +1(800)632-7799 email: [email protected]
External link: https://www.neb.com/protocols/2019/03/19/thermolabile-proteinase-k-typical-reaction-protocol
Protocol Citation: New England Biolabs 2022. Thermolabile Proteinase K Typical Reaction Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.bg9wjz7eVersion created by New England Biolabs
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it’s working
Created: June 07, 2020
Last Modified: February 10, 2022
Protocol Integer ID: 37910
Keywords: Proteinase K, serine protease, enzymatic reaction, related serine protease, dnase, rnase, peptide bond, detectable endonuclease, following incubation, variety of peptide bond,
Abstract
Thermolabile Proteinase K is an engineered, subtilisin-related serine protease that will hydrolyze a variety of peptide bonds and is frequently used to cleanup enzymatic reactions or cell lysates.
- Heat inactivated following incubation at 55°C for 10 minutes.
- Optimal activity and stability for up to 24 months.
- Active in a wide range of reaction buffers with optimal activity between 20 - 40°C and pH 7.0 - 9.5.
- No detectable endonuclease, exonuclease, DNase or RNase contaminating activities.
Materials
MATERIALS
Thermolabile Proteinase KNew England BiolabsCatalog #P8111S
Safety warnings
Please see SDS (Safety Data Sheet) for hazards and safety warnings.
Reactions may be scaled-up linearly to accommodate larger amounts of substrate and larger reaction volumes. Optimal buffering reagents, enzyme quantity, incubation temperatures and times may vary for particular substrates. Typical restriction enzyme cleanup conditions are as follows:
To a 50 µL restriction enzyme digest containing 1 µg DNA and 10 units of a restriction enzyme:
Add 1 µL Thermolabile Proteinase K , mix gently.
Incubate reaction at 37 °C for 00:15:00 .
Incubate reaction at 55 °C for 00:10:00 to inactivate Thermolabile Proteinase K.
Note
Note: Optimal reaction buffers, reaction temperatures as well as additional reaction properties of Thermolabile Proteinase K can be found in detail in the FAQ. Optimization may be required for other cleanup applications (ligation, extension, PCR, etc.)