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Thermolabile Proteinase K

Catalog # Concentration Size List Price Quantity Your Price
P8111S 120 units/ml 30 units $257.00
$231.30
Catalog # Size List Price Your Price
P8111S 30 units $257.00
$231.30
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

This recombinant, heat-inactivated Proteinase K is engineered for complete inactivation at 55°C in 10 minutes, with broad protease activity for digestion of proteins in nucleic acid preparations and other applications.

  • A unique, recombinant Proteinase K that retains similar activity and specificity as wild type but can be completely heat inactivated at lower temperatures compared to others.
  • Engineered for complete heat inactivation with incubation at 55°C for only 10 minutes.
  • No detectable endonuclease, exonuclease, DNase or RNase contaminating activities.
  • 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.
  • Broad spectrum subtilisin-related serine protease that hydrolyzes a variety of peptide bonds and is frequently used to cleanup enzymatic reactions or cell lysates.

Thermolabile Proteinase K is an engineered, subtilisin-related serine protease that will hydrolyze a variety of peptide bonds. It preferentially cleaves the peptide bond at the carboxyl side of aliphatic or aromatic amino acid residues. However; the specificity of Thermolabile Proteinase K can be broad. 

Thermolabile Proteinase K (TLPK) can be completely inactivated by incubation at 55°C for 10 minutes, which allows for subsequent enzymatic steps in the same reaction vessel. Figure 1 shows that the activity of restriction endonucleases, including heat-stable endonucleases, can be completely abolished using TLPK. 

 

Figure 1: Thermolabile Proteinase K completely degrades restriction enzyme activity



Lane L: 1 kb DNA Ladder (NEB #N3232); Lane 1: λ DNA incubated for 1 hour at 37°C; Lane 2: λ DNA incubated with PvuII-HF or PstI-HF for 1 hour at 37°C; Lane 3: λ DNA incubated with PvuII-HF or PstI-HF which had been treated with 1 μL Thermolabile Proteinase K (TLPK) for 10 minutes at 37 °C; Lane 4: λ DNA incubated first with TLPK treated PvuII-HF or PstI-HF followed by incubation at 55 °C for 10 minutes to inactivate the TLPK, followed by treatment with PvuII-HF or PstI-HF for 1 hour at 37°C. Results indicate that Thermolabile Proteinase K completely degrades restriction enzyme activity, allowing for subsequent enzymatic steps in the same reaction vessel.
Product Source
Cloned from Engyodontium album (formerly Tritirachium album), mutagenized to increase thermolability of the enzyme and expressed in Pichia pastoris.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Thermolabile Proteinase K P8111SVIAL -20 1 x 0.25 ml 120 units/ml
Features
  • Heat inactivated following incubation at 55°C for 10 minutes.
  • Isolation of plasmid and genomic DNA and RNA
  • Inactivation of RNases, DNases and various other enzymes in reaction workflows
  • PCR purification
An exception occurred during the operation, making the result invalid. Check InnerException for exception details.

Companion Products
Notes
  • Active in a wide range of buffers. It is highly active between pH 7.0 and 9.5 and temperatures 20-40°C. It is active in chelating agents such as EDTA up to 10 mM.
  • Thermolabile Proteinase K is stable for at least 2 years at –20°C. No loss of activity is observed after 10 freeze-thaw cycles.
  • Thermolabile Proteinase K is highly active over a broad pH range (7.0 - 9.5) and a wide temperature range (20 – 40°C).
  • Thermolabile Proteinase K is active in chelating agents such as EDTA up to 10 mM.
  • Enzyme activity is stimulated in up to 1% SDS and no inhibition is observed in SDS concentrations up to 2.5% or Triton X-100 concentrations up to 1.5%.
  • Thermolabile Proteinase K activity is inhibited by urea concentrations greater than 2 M.
Selection Charts
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]
Safety Data Sheets
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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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