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Proteinase K, recombinant, PCR Grade

from Titrirachium album, expressed in Pichia pastoris, lyophilizate

For further processing only.

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Order information
Proteinase K, recombinant, PCR Grade material number and pack size:
Material Number Pack Size
03508811103 25 mg
05963117103 5 g
03508838103 100 mg
03450376103 50 mg
05963133103 1 g
03450384103 250 mg
03450376103: Will be supplied as "Protein. K, rec PCR Grade Lyo. MPB 25 mg". Unit of measure is "piece".
03508811103: Will be supplied as "Protein. K, rec PCR Grade Lyo. MPB 50 mg". Unit of measure is "piece".
03508838103: Will be supplied as "Protein. K, rec PCR Grade Lyo. MPB 100 mg". Unit of measure is "piece".
03450384103: Will be supplied as "Protein. K, rec PCR Grade Lyo. MPB 250 mg". Unit of measure is "piece".
05963133103: Will be supplied as "Proteinase K, rec., PCR Grade, Lyo 1 g". Unit of measure is "piece".
05963117103: Will be supplied as "Proteinase K, rec., PCR Grade, Lyo 5 g". Unit of measure is "piece".
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Overview
Recombinant Proteinase K in PCR Grade quality is a universal tool for nucleic acid template preparation.
Proteinase K, originally isolated from the mold Tritirachium album, is a recombinant enzyme expressed in Pichia pastoris. It is a highly active, subtilisin-related serine endopeptidases that does not exhibit any pronounced cleavage specificity. Thus, Proteinase K, recombinant, PCR Grade, is a universal tool for template preparation. Amino acid sequence and molecular structure of the recombinant enzyme and the native protease are identical. However, the production process of the recombinant Proteinase K guarantees an enzyme of outstanding reliability and purity meeting all requirements of diagnostics' manufacturers.
Special emphasis has been placed on a low DNA-content of the enzyme preparation, making Proteinase K, recombinant, PCR Grade, ideally suited for isolating PCR and RT-PCR templates.
  • Maximize the yield of target nucleic acids.
    Proteinase K is rigorously tested for the absence of nucleases.
  • Experience stability and flexibility.
    The lyophilizate is very stable and can be stored at room temperature for at least 12 months. A broad range of fill volumes from 25 mg to 5 g per vial is available. Lyophilizate dissolved in water is stable for at least 60 days when stored at +2 to +4°C.
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Application
Proteinase K, recombinant, PCR Grade, digests native proteins very efficiently. This enzyme can be used to rapidly inactivate endogenous RNases and DNases during nucleic acid isolation. Proteinase K is particularly suited for the isolation of native RNA and DNA from tissues and cell lines.
The enzyme promotes cell lysis by activating a bacterial autolytic factor. Proteinase K is also used for:
  • Analysis of membrane structures by modifying proteins and glycoproteins on cell surfaces
  • Removal of cellular debris during the preparation of colony lifts
  • Treatment of tissue sections to ensure efficient probe infiltration during in situ hybridization
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Specification
Nomenclature: Proteinase K
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 and does not exhibit any pronounced cleavage specificity. Activity can be stimulated by addition of denaturing agents (SDS and urea).
Appearance: White lyophilizate
Solubility: Clear, colorless solution in water (c=20 mg/mL)
Volume activity (+37°C, hemoglobin): ≥24 U/mg lyophilizate
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.
Volume activity (+25°C, Chromozym): ≥2 U/mg lyophilizate
Specific activity (+25°C, Chromozym): ≥2.5 U/mg protein
Unit definition (Chromozym): One unit is the enzyme activity which cleaves 8 mmol Chromozym TRY in 1 minute at +25°C.
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: ≤125 CFU/g
Stability: At +2 to +8°C within specification range for 18 months.