Proteinase K, rec., PCR Grade, sol.

Maximize the yield of target nucleic acids.

Proteinase K is rigorously tested for the absence of nucleases.

Experience stability and safety.

The solution is very stable and can be stored at room temperature for at least 18 months. The handling of the solution is safe and flexible.

Application

Proteinase K, recombinant, PCR Grade, digests proteins very efficiently. It can be used to rapidly inactivate endogenous RNases and DNases during nucleic acid isolation or remove enzymes that have been used during (m)RNA synthesis by in vitro transcription, as in the manufacture of mRNA vaccines.

Proteinase K is also used to:

Isolate native RNA and DNA from tissues and cell lines by promoting cell lysis

Analyze membrane structures by modifying proteins and glycoproteins on cell surfaces

Remove cellular debris during the preparation of colony lifts

Treat tissue sections to ensure efficient probe infiltration during in situ hybridization

Properties

Proteinase K, originally isolated from the mold Tritirachium album, is a recombinant enzyme expressed in Pichia pastoris.It is a highly active, unspecific subtilisin-related serine endopeptidase.

EC Number: 3.4.21.64

CAS Number: 39450-01-6

Molecular weight: 28.8 kD

pH optimum: 7.5-10.5

Inhibitors: Proteinase K is inhibited by diisopropyl fluorophosphate and phenylmethylsulfonyl fluoride (PMSF) and is also totally inactivated by mercuric ions. Pefabloc® SC and Pefabloc® PLUS are specific, irreversible and nontoxic inhibitors.

Specificity: Proteinase K is one of the most active endopeptidases known without any pronounced cleavage specificity. Activity can be stimulated by addition of denaturing agents (SDS and urea).

Stability: Even at room temperature, the solution is very stable and can be stored for at least 18 months.

Specifications

Appearance: Clear, colorless solution

Volume activity (+37°C, hemoglobin): ≥600 U/mL

Specific activity (+37°C, hemoglobin): ≥30 U/mg protein

Unit definition (hemoglobin): One unit is the enzyme activity which releases folin positive amino acids and peptides equivalent to 1 μmol of tyrosine in 1 minute under the test conditions at 37°C.

Volume activity (+25°C, Chromozym): ≥50 U/mL

Specific activity (+25°C, Chromozym): ≥2.5 U/mg protein

Unit definition (Chromozym): One unit is the enzyme activity which cleaves 18 mmol Chromozym TRY in 1 minute at +25°C.

Protein (Biuret): 14.0-22.0 mg/mL

Unspecific endonucleases (MWM III DNA): Not detectable in up to 200 μg after 16 hours incubation at +37°C.

Nicking activity (pBR322 DNA): Not detectable in up to 200 μg after 16 hours incubation at +37°C.

Ribonucleases (MS2 RNA): Not detectable in up to 40 μg after 16 hours incubation at +37°C.

DNA (Threshold®): ≤10 pg/mg enzyme

Bioburden: ≤5 CFU/mL

Stability: At +2 to +8°C within specification range for 18 months.

Regulatory Disclaimer

For further processing only.