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

03-32-1501
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      Overview

      Replacement Information

      Key Specifications Table

      Pricing & Availability

      Catalog NumberAvailability Packaging Qty/Pack Price Quantity
      03-32-1501-25MG
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          Plastic ampoule 25 mg
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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
          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

          SDS

          Title

          Safety Data Sheet (SDS) 

          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.