03-32-1501 Z-Phe-Arg-7-amido-4-methylcoumarin, Hydrochloride - CAS 70382-26-2 - Calbiochem

03-32-1501
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03-32-1501-25MG
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      Description
      OverviewSubstrate for fluorogenic assay of plasma and glandular kallikreins. Also a substrate for cathepsin B, cathepsin L, and papain.
      Catalogue Number03-32-1501
      Brand Family Calbiochem®
      References
      ReferencesMorita, T., et al. 1977. J. Biochem. 82, 1495.
      Product Information
      CAS number70382-26-2
      ATP CompetitiveN
      FormWhite to off-white solid
      Hill FormulaC₃₃H₃₆N₆O₆·HCl
      Chemical formulaC₃₃H₃₆N₆O₆·HCl
      ReversibleN
      Quality LevelMQ200
      Applications
      Biological Information
      Primary TargetSubstrate for fluorogenic assay of plasma and glandular kallikreins
      Purity≥98% by HPLC
      Physicochemical Information
      Cell permeableN
      Peptide SequenceZ-Phe-Arg-AMC, HCl
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      R PhraseR: 36/37/38

      Irritating to eyes, respiratory system and skin.
      S PhraseS: 26-36

      In case of contact with eyes, rinse immediately with plenty of water and seek medical advice.
      Wear suitable protective clothing.
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Ambient Temperature Only
      Toxicity Irritant
      Storage -20°C
      Protect from Moisture Protect from moisture
      Do not freeze Ok to freeze
      Special InstructionsFollowing reconstitution, aliquot and freeze (-20°C). Stock solutions are stable for up to 3 months at -20°C.
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      Z-Phe-Arg-7-amido-4-methylcoumarin, Hydrochloride - CAS 70382-26-2 - Calbiochem MSDS

      Title

      Safety Data Sheet (SDS) 

      Z-Phe-Arg-7-amido-4-methylcoumarin, Hydrochloride - CAS 70382-26-2 - Calbiochem Certificates of Analysis

      TitleLot Number
      03-32-1501

      References

      Reference overview
      Morita, T., et al. 1977. J. Biochem. 82, 1495.

      Brochure

      Title
      Cathepsins and Related Products Technical Bulletin
      Data Sheet

      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.

      Revision18-September-2008 RFH
      DescriptionSubstrate for fluorogenic assay of plasma and glandular kallikreins. Also serves as a substrate for cathepsin B, cathepsin L, and papain.
      FormWhite to off-white solid
      Recommended reaction conditions
      Protocol for the Assay of Cathepsin L Enzyme: Cathepsin L is one of the lysosomal cysteine proteinases involved in the turnover of endocytosed proteins. It also has a higher specific activity towards physiological substrates, such as collagen and fibronectin over other lysosomal enzymes. It has an acidic optimal pH of 5.5. It is rapidly inactivated, 85% loss of activity in ~15 min, at pH 7.0-7.4. Substrate: Z-Phe-Arg-7-amido-4-methylcoumarin is more sensitive to cathepsin L than cathepsin B, although both enzymes will cleave the substrate. To determine cathepsin L activity, the specific cathepsin B substrate, Z-Arg-Arg-7-amido-4-methylcoumarin (Cat. No. 03-32-1570), can be used. The substrate is non-fluorescent, but when hydrolyzed by the enzyme it releases highly fluorescent 7-amido-4-methylcoumarin (AMC). AMC can be quantitated with a spectrofluorometer using an excitation at ~370 nm and an emission at ~460 nm. Reagents: • Assay/Activator: 340 mM sodium acetate, 60 mM acetic acid, and 4 mM EDTA, Buffer pH 5.5; add DTT to a final concentration of 8 mM just prior to use. • Substrate: Prepare a 1 mM Z-Phe-Arg-7-amido-4-methylcoumarin stock solution in DMSO and store in a -20°C freezer. Dilute to 20 µM with H2O prior to use. • Stop Solution: 100 mM sodium monochloroacetate, 70 mM acetic acid, and 30 mM sodium acetate, pH 4.3 • Diluent: BRIJ® 35 (0.1% in H2O) • Standard: Prepare a 1 mM AMC stock solution in DMSO and store in a 4°C refrigerator. Dilutions are made using a 1:1 mixture of Assay Buffer and Stop Solution. Procedure: 1. Using the Diluent, prepare 500 µl of enzyme sample in 5 ml glass or polystyrene tubes. 2. Add 250 µl of assay/activator buffer. Allow ~1 min. incubation at 30°C for temperature equilibration and enzyme activation. Note: longer incubation may result in loss of activity. 3. Add 250 µl of the 20 µM substrate solution and mix. 4. Following a 10 min incubation at 30°C, add 1 ml of Stop Solution. 5. Measure the fluorescence and determine the molar amount using a standard curve for AMC. The fluorescence of AMC is only minimally affected by pH in the range of 4-7.
      CAS number70382-26-2
      Chemical formulaC₃₃H₃₆N₆O₆·HCl
      Peptide SequenceZ-Phe-Arg-AMC, HCl
      Purity≥98% by HPLC
      Solubility5% Acetic acid (1 mg/ml), H₂O (1 mg/ml), or DMSO (1 mg/ml)
      Storage -20°C
      Protect from moisture
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing reconstitution, aliquot and freeze (-20°C). Stock solutions are stable for up to 3 months at -20°C.
      Toxicity Irritant
      ReferencesMorita, T., et al. 1977. J. Biochem. 82, 1495.