Ordering Information
- : 539480-100MG
- : 100 MG
Proteinase K, Tritirachium album
Description:
Proteinase K, Tritirachium album
Trade Name:
Calbiochem (EMD Millipore)
Qty/Pk:
100 MG
Background Information:
Serine protease that exhibits strong proteolytic activity on a wide variety of denatured and native proteins of high molecular weight. Because of its rapid proteolytic inactivation of endogenous nucleases, it can be used for isolation of mRNA and high molecular weight DNA. Preferentially cleaves bonds next to the carbonyl group of N-substituted hydrophobic, aliphatic, and aromatic amino acids. Inhibited by DFP, Hg2+, and PMSF. Not inactivated by metal chelators, sulfhydryl reagents, TLCK, or TPCK.
Application Notes:
Anson Enzyme Activity Assay:
Materials Required
• 5 ml of Hemoglobin (20 mg/ml) in 100 mM phosphate buffer, pH 7.5 (6 M urea, 5 mM NaCl).
• 0.6 ml of 20 mM CaCl2 dissolved in distilled water.
• 0.1 ml of proteinase K (40 µg/ml) in 20 mM CaCl2
Activity Assay Protocol
1. Mix the 5 ml hemoglobin, 0.6 ml CaCl2, and 0.1 ml proteinase K and make up the volume to 6 ml with 20 mM CaCl2; incubate for 10 min at 25°C.
2. Add 10 ml 300 mM Trichloroacetic acid and incubate at room temperature for 30 min.
3. Centrifuge and transfer 5 ml of the supernatant to a clean tube. Add 10 ml 500 mM NaOH and 3 ml Folin reagent (diluted 1:2 with redistilled water); incubate for 5 min at room temperature.
4. Measure the ΔA at 750 nm. For calibration use tyrosine solutions. 1 mAnson unit liberates 1 µmole of Folin-positive amino acids per min (calculated as tyrosine) under test conditions.
Materials Required
• 5 ml of Hemoglobin (20 mg/ml) in 100 mM phosphate buffer, pH 7.5 (6 M urea, 5 mM NaCl).
• 0.6 ml of 20 mM CaCl2 dissolved in distilled water.
• 0.1 ml of proteinase K (40 µg/ml) in 20 mM CaCl2
Activity Assay Protocol
1. Mix the 5 ml hemoglobin, 0.6 ml CaCl2, and 0.1 ml proteinase K and make up the volume to 6 ml with 20 mM CaCl2; incubate for 10 min at 25°C.
2. Add 10 ml 300 mM Trichloroacetic acid and incubate at room temperature for 30 min.
3. Centrifuge and transfer 5 ml of the supernatant to a clean tube. Add 10 ml 500 mM NaOH and 3 ml Folin reagent (diluted 1:2 with redistilled water); incubate for 5 min at room temperature.
4. Measure the ΔA at 750 nm. For calibration use tyrosine solutions. 1 mAnson unit liberates 1 µmole of Folin-positive amino acids per min (calculated as tyrosine) under test conditions.
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Molecular Weight:
28500
CAS No.:
39450-01-6
Brand Family:
Calbiochem
Presentation:
White lyophilized solid. Lyophilized from calcium acetate
Contaminants: DNase: no detectable nicking activity with pBR322, incubation for 6 h at 37°C; RNase: no ribonuclease activity with MS2 RNA detected after incubation for 6 h at 37°C
Contaminants: DNase: no detectable nicking activity with pBR322, incubation for 6 h at 37°C; RNase: no ribonuclease activity with MS2 RNA detected after incubation for 6 h at 37°C
Storage Conditions:
Shipped at Ambient Temperature. Store at +2 - 8°C.
Specific Activity:
≥40 mAnsonU/mg protein
Properties:
Solubility: 50 mM Tris-HCl, 2 mM calcium acetate, pH 8.0 .
Unit Definition:
One unit is defined as the amount of enzyme that will liberate 1.0 µmol tyrosine from casein per min at 37°C, pH 7.5.
Product Name:
Proteinase K, Tritirachium album
Activity:
≥30 mAnsonU/mg dry weight