Description
QIAGEN Protease and QIAGEN Proteinase K offer broad substrate specificity with high activity for a wide range of reaction conditions. QIAGEN Proteinase K in 2 mL and 10 mL pack sizes is dedicated for use in various QIAGEN kits (please see list in the Principle section below). As a sample prep expert, QIAGEN also offers Proteinase K as lyophilized powder (Proteinase K Powder). Both proteases offer high activity in buffers commonly used in most DNA and RNA isolation procedures and are quality-guaranteed by QIAGEN.
QIAGEN Protease and QIAGEN Proteinase K QIAGEN Protease and QIAGEN Proteinase K offer broad substrate specificity with high activity in buffers commonly used in most DNA and RNA isolation procedures. Both enzymes are quality-guaranteed by QIAGEN. QIAGEN Protease is particularly stable and active at high pH. In Tris-containing buffers of alkaline pH (7.5–10.5) QIAGEN Protease displays 30 to 40% higher specific activity than Proteinase K. In the presence of strong denaturants, such as urea (0.75–3 M urea) or guanidine HCl (0.5–3.0 GuHCl), the specific activity is comparable to that of Proteinase K, provided the EDTA concentration is less than 8 mM. QIAGEN Protease shows increased activity in the presence of urea and guanidine HCl. Similar stimulation is obtained upon the addition of up to 1% SDS. Enzymatic activity also increases as a function of temperature (30–55°C). For more detailed information, refer to the QIAGEN Protease Product Sheet in the Resources section. The enzyme can be easily inactivated by incubation at 70°C for 15 minutes. QIAGEN Protease is not inhibited by up to 100 mM EDTA in Tris·Cl buffers. However, in the presence of greater than 1% SDS or other strong denaturants, the EDTA concentration must be less than 8 mM. Proteinase K powder Proteinase K powder is a lyophilized, white powder, highly active with exceptional lot-to-lot consistency (see figure “High lot-to-lot consistency Proteinase K powder”), ensuring reproducibility for stable working conditions, repeatable and reliable experiment results. Proteinase K powder is highly stable when stored at –20°C (see figure “Proteinase K powder stability”). Proteinase K powder specifications: No detected exonuclease, endonuclease or RNase activity Solubility in water ≥20 mg/mL; activity ≥30 U/mg lyophilizate; specific activity ≥40 U/mg protein; protein content ≥70%; DNA content ≤10 pg/mg Shipping conditions: Ambient temperature
Technical specifications QIAGEN Protease QIAGEN Proteinase K Proteinase K Powder Format Lyophilized powder Ready-to-use solution Lyophilized powder Amount 7.5 AU or 4 x 7.5 AU 2 mL or 10 mL (20 mg/mL) 20 mg, 100 mg, 250 mg, 1 g, 10 g, 25 g, 50 g, 200 g Activity 45 mAU/mg protein >600 mAU/mL ≥30 U/mg lyophilizate ≥40 U/mg protein Unit definition One mAU is the activity that releases folin-positive amino acids and peptides corresponding to 1 µmol tyrosine per minute One unit of Proteinase K hydrolyzes urea-denatured hemoglobin producing the color equivalent of 1 μ mol tyrosine per 1 minute at 37°C and pH 7.5 (Folin & Ciocalteu’s method), 1 U = 1 mAnsonU QIAGEN Proteinase K (in 2 mL and 10 mL pack sizes) is a subtilisin-type protease isolated from the saprophytic fungus Tritirachium album and is particularly suitable for short digestion times. It possesses a high specific activity that remains stable over a wide range of temperatures and pH values with substantially increased activity at a higher temperature. Soluble calcium is not essential for enzymatic activity. This means that EDTA, which is used to inhibit Mg 2+ -dependent enzymes, such as nucleases, will not inhibit Proteinase K activity. Proteinase K is supplied in the following QIAGEN kits: QIAamp Circulating Nucleic Acid Kit QIAamp DNA Mini Kit QIAamp Fast DNA Stool Mini Kit DNeasy Blood & Tissue Kit DNeasy 96 Blood & Tissue Kit QIAamp DNA Micro Kit QIAamp MinElute Media Kit QIAamp Media MDx Kit QIAGEN Protease is a serine protease isolated from a recombinant Bacillus strain. It is an economical alternative to Proteinase K for isolation of native DNA and RNA from a variety of sources. QIAGEN Protease is completely free of DNase and RNase activities. QIAGEN Protease is supplied in the following QIAGEN kits: QIAamp DNA Blood Mini, Midi and Maxi Kits QIAamp 96 DNA Blood Kit Blood & Cell Culture DNA Kits QIAamp Virus BioRobot 9604 Kit QIAamp DNA Blood BioRobot MDx Kit QIAamp DSP Virus Kit QIAamp DSP DNA Blood Mini Kit Note: Users of manual QIAamp DNA Blood Kits and QIAGEN Blood & Cell Culture DNA Kits should resuspend each bottle of QIAGEN Protease with 7 mL distilled water. Note: Users of the QIAamp DNA Blood BioRobot 9604 Kit should resuspend each bottle of QIAGEN Protease with 10 mL distilled water. Note: QIAGEN Protease is not compatible with Buffer ATL in the DNeasy Tissue, DNeasy 96 Tissue and QIAamp DNA Mini Kit. In the presence of >0.5% SDS, >1% sarkosyl or high concentrations of other detergents, the EDTA concentration must be <8 mM for full activity over extended incubation times.
Instructions for using QIAGEN Protease, QIAGEN Proteinase K and Proteinase K Powder are provided in the corresponding kit handbook.
