Highly characterized for more consistent performance, Proteinase K is a subtilisin-relatedserine protease that will hydrolyze a variety ofpeptide bonds. Proteinase K is active in a widerange of temperatures and buffers with optimalactivity between 20 and 60°C and a pH between 7.5and 12.0 (1, 2). Activity is stimulated when up to2% SDS or up to 4 M urea are included in the reaction(3). Calcium is important for thermostabilityof Proteinase K but it is not required for catalysis,therefore Proteinase K is also active in bufferscontaining chelating agents such as EDTA (4).
Protein Concentration
20 mg/ml, approximately, as determined by UV absorption at 280 nm.
Product Source
Engyodontium album (Tritirachium album)Reagents Supplied
The following reagents are supplied with this product:
| NEB # | Component Name | Component # | Stored at (°C) | Amount | Concentration |
| P8107S | -20 |
| Proteinase K, Molecular Biology Grade | P8107SVIAL | -20 | 2 x 1 ml | 800 units/ml |
Application Features
- Isolation of plasmid and genomic DNA
- Isolation of RNA
- Inactivation of RNases, DNases and enzymes in reactions
- Removal of enzymes from DNA to improve cloning efficiency (5)
- PCR purification
Properties & Usage
Unit Definition
One unit will digest urea-denatured hemoglobin at 37°C (pH 7.5) per minute to produce equal absorbance as 1.0 μmol of L-tyrosine using Folin & Ciocalteu's phenol reagent (6).
Storage Buffer
20 mM Tris-HCl
1 mM CaCl2
50% Glycerol
pH 7.4 @ 25°C
Heat Inactivation
95°C for 10 minutes
Molecular Weight
Theoretical: 28.9 kDa
Unit Assay Conditions
0.5–2 μg of Proteinase K is incubated with 2% denatured hemoglobin solution for 10 minutes at 37°C (pH 7.5). After precipitation, neutralization and addition of Folin & Ciocalteu's phenol reagent, absorbance of soluble cleavage products are measured at 750 nm. Absorbance is compared to a standard curve of L-tyrosine absorbance prepared similarly.
Notes
- Active in a wide range of buffers, including all NEB-specific restriction endonuclease buffers. It is highly active between pH 7.5 and 12 and temperatures 20-60°C. Proteinase K is also active in chelating agents such as EDTA and activity is stimulated in up to 2% SDS or 4M urea.
- Proteinase K is stable for at least 2 years at –20°C. No loss of activity is observed after 10 freeze-thaw cycles.
- Reaction Conditions
Proteinase K is active in a wide range of buffers including all NEB specific restriction endonuclease buffers. It is highly active between pH 7.5 and 12.0 and temperatures between 20 and 60°C (1,2). Proteinase K is also active in chelating agents such as EDTA (4) and activity is stimulated in up to 2% SDS or 4 M urea (3).
References
- Bajorath, J. et al. (1988). Biochimica et Biophysica Acta. 954, 176-182.
- Ebeling, W. et al. (1974). Eur. J. Biochem. 47, 91-97.
- Hilz, H. et al. (1975). Eur. J. Biochem. 56, 103-108.
- Bajorath, J. et al. (1988). Eur. J. Biochem. 176, 441-447.
- Crowe, J.S. et al, (1991). Nucleic Acids Research. 91, 184.
- Anson, M.L. (1939). J. Gen. Physiol. 22, 79-89.
- Pace, C.N. et al. (1995). Protein Sci. 4, 2411-2423.
Protocols
- General protocol to release nucleic acids prior to capillary or gel electrophoresis using Proteinase K (P8107)
- Protocol to cleanup DNA Glucosylation/restriction digest and Proteinase using Proteinase K (P8107)
- Protocol to purify PCR products in preparation for cloning using Proteinase K (P8107)
- Using recombinant Cas9 nuclease to assess locus modification in genome editing experiments (#M0386)
FAQs
- The protocol I am using requires a µg/ml concentration of Proteinase K. What is the protein concentration of Proteinase K, Molecular Biology Grade?
- What buffer should I use for Proteinase K?
- If I need to dilute Proteinase K, Molecular Biology Grade, how should I do this?
- Why is the unit definition assay for Thermolabile Proteinase K (NEB #P8111) different from the unit definition assay for Proteinase K, Molecular Biology Grade (NEB # P8107)?
- What is the activity if Proteinase K, Molecular Biology Grade if subjected to the unit definition assay used for Thermolabile Proteinase K?
Citations
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
- P8107S_v1_0171803
- P8107S_v1_10013526
- P8107S_v1_10016569
- P8107S_v1_10018079
- P8107S_v1_10021955
- P8107S_v1_10025786
- P8107S_v1_10028131
- P8107S_v1_10033007
- P8107S_v1_10035054
- P8107S_v1_10039754
- P8107S_v1_10042777
- P8107S_v1_10046009
- P8107S_v1_10047990
- P8107S_v1_10050147
- P8107S_v1_10052521
- P8107S_v1_10054881
- P8107S_v1_10058651
- P8107S_v1_10060508
- P8107S_v1_10061037
- P8107S_v1_10065104
- P8107S_v1_10067123
- P8107S_v1_10072120
- P8107S_v2_10081061
- P8107S_v2_10081790
- P8107S_v2_10082241
- P8107S_v2_10083332
- P8107S_v2_10085165
- P8107S_v2_10086180
- P8107S_v2_10087434
- P8107S_v2_10089091
- P8107S_v2_10090445
- P8107S_v2_10092744
- P8107S_v2_10093426
- P8107S_v2_10094130
- P8107S_v2_10096309
- P8107S_v2_10100138
- P8107S_v2_10106667
- P8107S_v2_10107398
- P8107S_v2_10112434
- P8107S_v2_10117872
- P8107S_v2_10121119
- P8107S_v2_10127339
- P8107S_v2_10130705
- P8107S_v2_10133499
- P8107S_v2_10144695
- P8107S_v2_10145593
- P8107S_v2_10156147
- P8107S_v2_10164847
- P8107S_v2_10175667
- P8107S_v2_10179177
- P8107S_v2_10185534
- P8107S_v2_10190420
- P8107S_v2_10195886
- P8107S_v2_10204667
- P8107S_v2_10213055
- P8107S_v2_10216318
- P8107S_v2_10221126
- P8107S_v2_10229227
- P8107S_v2_10235815
- P8107S_v2_10241266
- P8107S_v2_10250777
Safety Data Sheets
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