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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.
Catalogue Number
539480
Brand Family
Calbiochem®
References
References
Watazu, Y., et al. 1993. J. Lab. Clin. Anal. 7, 81. Lebherz, H.G., et al. 1986. Biochem. J.233, 51. Ebeling, W., et al. 1974. Eur. J. Biochem.47, 91.
Product Information
CAS number
39450-01-6
Activity
≥30 mAnsonU/mg dry weight
Unit of Definition
One mAnson unit is defined as the amount of enzyme that will liberate 1.0 µmol of Folin-positive amino acids (calculated as tyrosine) per min at 37°C, pH 7.5.
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 and Shipping Information
Ship Code
Ambient Temperature Only
Toxicity
Irritant
Storage
+2°C to +8°C
Do not freeze
Ok to freeze
Special Instructions
Following reconstitution, store in the refrigerator (4°C). Stock solutions prepared in 50 mM Tris-HCl, 2 mM calcium acetate, pH 8.0 are stable for up to 1 year at 4°C.
Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.
Revision
16-November-2016 JSW
Description
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.
Form
White lyophilized solid
Formulation
Lyophilized from calcium acetate
Recommended reaction conditions
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 CaCl2Activity 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.
CAS number
39450-01-6
EC number
3.4.21.14
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
Specific activity
≥40 mAnsonU/mg protein
Activity
≥30 mAnsonU/mg dry weight
Unit definition
One mAnson unit is defined as the amount of enzyme that will liberate 1.0 µmol of Folin-positive amino acids (calculated as tyrosine) per min at 37°C, pH 7.5.
Solubility
50 mM Tris-HCl, 2 mM calcium acetate, pH 8.0
Storage
+2°C to +8°C
Do Not Freeze
Ok to freeze
Special Instructions
Following reconstitution, store in the refrigerator (4°C). Stock solutions prepared in 50 mM Tris-HCl, 2 mM calcium acetate, pH 8.0 are stable for up to 1 year at 4°C.
Toxicity
Irritant
References
Watazu, Y., et al. 1993. J. Lab. Clin. Anal. 7, 81. Lebherz, H.G., et al. 1986. Biochem. J.233, 51. Ebeling, W., et al. 1974. Eur. J. Biochem.47, 91.
