Peptidase and Protease Substrates

• AMC- or MCA-based Substrates
  • 7-Amino-4-methylcoumarin, N-CBZ-L-isoleucyl-L-glutamyl-L-threonyl-L-aspartic acid amide (Z-IETD-AMC, or Z-IETD-MCA))

    Z-IETD-AMC is a substrate for caspase-8, which is involved at the early stage of apoptosis, acting as an initiator of the caspase activation cascade.

    • Catalog Number: 10200
    • Absorption/emission of substrate = 330/390nm (weak fluorescence);
    • Absorption/emission of end product (AMC) = 342/441nm (strong fluorescence)
    • Off-white solid soluble in DMSO.
    • C₃₇H₄₅N₅O₁₃.
    • Mwt: 767.79.
    
    • References
      1. J Cell Biol 155, 637(2001);
      2. J Biol Chem 274, 23426(1999); 3) J Biol Chem 274, 22686(1999).
  • 7-Amino-4-methylcoumarin, N-CBZ-L-phenylalanyl-L-arginine amide, hydrochloride (Z-FR-AMC, or Z-FR-MCA)

    Z-FR-AMC is a substrate for serine proteases, including cathepsins, kallikrein and plasmin.

    • Catalog Number: 10201
    • Absorption/emission of substrate = 330/390nm (weak fluorescence); absorption/emission of end product (AMC) = 342/441nm (strong fluorescence).
    • Off-white solid soluble in DMSO.
    • Store at -20℃ and protect from light.
    • C₃₃H₃₇ClN₆O₆.
    • Mwt: 649.14.
    
    
    • References
      1. J Biol Chem 270, 558(1995);
      2. Arch Biochem Biophys 314, 171(1994);
      3. FEBS Lett 341, 197(1994);
      4. J Biochem (Tokyo) 113, 441(1993);
      5. Biochem J 201, 367(1982).
  • 7-Amino-4-methylcoumarin, N-CBZ-L-aspartyl-L-glutamyl-L-valyl-l-aspartic acid amide (Z-DEVD-AMC, or Z-DEVD-MCA)

    Z-DEVD-AMC is a fluorogenic substrate for caspase 3 and can be used to continuously measure the activity of caspase-3 in cell extracts, using a fluorometer or fluorescence microplate reader.

    • Catalog Number: 10202
    • Absorption/emission of substrate = 330/390nm (weak fluorescence); absorption/emission of end product (AMC) = 342/441nm (strong fluorescence).
    • Off-white solid soluble in DMSO.
    • Store at -20℃ and protect from light.
    • C₃₆H₄₁N₅O₁₄.
    • Mwt: 767.74.
    
    
    • References
      1. Anal Biochem 292, 313 (2001);
      2. Biochemistry 39, 16056(2000):
      3. Br J Pharmacol 129, 1673(2000);
      4. J Biol Chem 275, 21094(2000).
• AFC-based Substrates
  • 7-Amino-4-trifluoromethylcoumarin, N-CBZ-L-aspartyl-L-glutamyl-L-valyl-l-aspartic acid amide (Z-DEVD-AFC)

    Z-DEVD-AFC is a fluorogenic substrate for caspase 3 and can be used to continuously measure the activity of caspase-3 in cell extracts, using a fluorometer or fluorescence microplate reader.

    • Catalog Number: 10203
    • Absorption/emission of substrate = 340/434nm (weak fluorescence); absorption/emission of end product (AFC) = 400/505nm (strong fluorescence).
    • Light yellow solid soluble in DMSO.
    • Store at -20℃ and protect from light.
    • C₃₆H₃₈F₃N₅O₁₄.
    • Mwt: 821.7.
    
    
    • References
      1. J Biol Chem 277, 804(2002);
      2. J Cell Biol 156, 1077(2002);
      3. Apoptosis 6, 151(2001);
      4. J Biol Chem 276, 34244(2001);
      5. J Neurosci 18, 3659(1998);
      6. Proc Natl Acad Sci U S A 93, 14559(1996).
  • 7-Amino-4-trifluoromethylcoumarin, N-CBZ-L-isoleucyl-L-glutamyl-L-threonyl-L-aspartic acid amide (Z-IETD-AFC)

    Z-IETD-AFC is a substrate for caspase-8, which is involved at the early stage of apoptosis, acting as an initiator of the caspase activation cascade. Absorption/emission of substrate = 340/434nm (weak fluorescence);

    • Catalog Number: 10204
    • absorption/emission of end product (AFC) = 400/505nm (strong fluorescence).
    • Light yellow solid soluble in DMSO.
    • Store at -20℃ and protect from light.
    • C₃₇H₄₂F₃N₅O₁₃.
    • Mwt: 821.76.
    
    
    • References
      1. J Biol Chem 276, 14545(2001);
      2. J Biol Chem 276, 27432(2001);
      3. J Immunol 166, 6473(2001);
      4. J Biol Chem 275, 11814(2000).
• Rhodamine 110-based Substrates
  • Rhodamine 110, bis-(L-aspartic acid amide), ditrifluoroacetic acid salt (D₂R)

    D₂R is a general substrate for caspases including caspases 3 and 7. Unlike other fluorogenic substrates for caspases, D₂R appears to be membrane-permeant, and thus can be used to measure caspase activity in live cells without osmotic shock for flow cytometry studies, as well as for cell extracts.

    • Catalog Number: 10210
    • Absorption/emission of end product (R110) = 496/520nm.
    • Off-white to pink solid soluble in DMSO.
    • Store at -20℃ and protect from light.
    • C₃₂H₂₆F₆N₄O₁₃.
    • Mwt: 788.57.
    
    
    • References
      1. Biochemistry 38, 13906(1999).
  • Rhodamine 110, bis-(N-CBZ-L-arginine amide), dihydrochloride (BZAR)

    BZAR is a substrate for serine proteases,^1-5 and is also an inhibitor for quanidinobenzoatase.⁶ It is 50 to 100 times more sensitive than the AMC-based substrate.

    • Catalog Number: 10205
    • Absorption/emission of end product (R110) = 496/520nm.
    • Off-white to pink solid soluble in DMSO.
    • Store at -20℃ and protect from light.
    • C₄₈H₅₂Cl₂N₁₀O₉
    • Mwt: 983.91.
    
    
    • References
      1. J Histochem Cytochem 49, 1473(2001);
      2. Biotechniques 29, 1108-1113 (2000);
      3. J Enzyme Inhib 6, 303(1992);
      4. Biochem Soc Trans 18, 632 (1990);
      5. J Cancer Res Clin Oncol 116, 57(1990);
      6. J Enzyme Inhib 2, 209(1988).
  • Rhodamine 110, bis-(N-CBZ-L-aspartyl-L-glutamyl-L-valyl-l-aspartic acid amide) (Z-DEVD-R110)

    Z-DEVD-R110 is a fluorogenic substrate for caspase 3 and can be used to continuously measure the activity of caspase-3 in cell extracts, using a fluorometer or fluorescence microplate reader. Absorption/emission of end product (R110) = 496/520nm (strong fluorescence). Of-white solid soluble in DMSO. Store at -20℃ and protect from light. C₇₂H₇₈N₁₀O₂₇. Mwt: 1514.

    • Catalog Number: 10206
    
    
    • References
      1. J Biol Chem 275, 288(2000);
      2. Biochemistry 38, 13906(1999).
  • Rhodamine 110, bis-(N-CBZ-L-isoleucyl-L-glutamyl-L-threonyl-L-aspartic acid amide) (Z-IETD-R110)

    Z-IETD-R110 is a substrate for caspase-8, which is involved at the early stage of apoptosis, acting as an initiator of the caspase activation cascade.

    • Catalog Number: 10207
    • Absorption/emission of end product (R110) = 496/520nm.
    • Off-white to pink solid soluble in DMSO.
    • Store at -20℃ and protect from light.
    • C₇₄H₈₆N₁₀O₂₅.
    • Mwt: 1514
    
    
  • Rhodamine 110, bis-(N-CBZ-L-isoleucyl-L-prolyl-L-arginine amide), dihydrochloride (BZiPAR)

    BZiPAR is a substrate for trypsin.^1,2 The substrate has been reported to enter live cells and be hydrolyzed by lysosomal proteases.³

    • Catalog Number: 10208
    • Absorption/emission of end product (R110) = 496/520nm.
    • Off-white to pink solid soluble in DMSO.
    • Store at -20℃ and protect from light.
    • C₇₀H₈₈Cl₂N₁₄O₁₃.
    • Mwt: 1404.5
    
    
    • References
      1. Proc Natl Acad Sci U S A 97, 13126(2000);
      2. Biochim Biophys Acta 788, 74(1984);
      3. Photochem Photobiol 44, 461(1986).
  • Rhodamine 110, bis-(N-CBZ-L-phenylalanyl-L-arginine amide), dihydrochloride (Z-FR-R110)

    Z-FR-R110 is a substrate for the cysteine proteases cathepsins B and L. The substrate has been used in flow cytometry studies of human monocytes and rat macrophages.

    • Catalog Number: 10209
    • Absorption/emission of end product (R110) = 496/520nm.
    • Off-white to pink solid soluble in DMSO.
    • Store at -20℃ and protect from light.
    • C₆₆H₇₂Cl₂N₁₂O₁₁.
    • Mwt: 1278.26
    
    
    • References
      1. Glia 7, 183(1993);
      2. Biol Chem Hoppe Seyler 373, 433(1992);
      3. Biol Chem Hoppe Seyler 373, 547(1992).

• Bioluminescent Substrates
• Phosphatase Substrates
• Substrates For Redox Enzymes
• Substrates For beta-galactosidase