Calpain-1 substrate, fluorogenic, is a sensitive and specific substrate for calpain-1 that cleaves the Tyr-Gly bond, resulting in enhanced fluorescence [1] (Ex/Em = 490 nm/518 nm).
Ac-DNLD-AMC is a fluorogenic caspase-3 substrate. Upon enzymatic cleavage by caspase-3, 7-amino-4-methylcoumarin (AMC) is released and its fluorescence can be used to quantify caspase-3 activity. AMC displays excitation/emission maxima of 340-360 and 440-460 nm, respectively.
STY-BODIPY (Styrene-BODIPY) is a styrene-conjugated fluorescent probe enabling calculation of radical trapping antioxidant (RTA) activity by monitoring absorbance loss at 571 nm.
Mca-PLAC(p-OMeBz)-WAR(Dpa)-NH2 is a fluorogenic substrate for matrix metalloproteinase-14 (MMP-14). Upon cleavage by MMP-14, 7-methoxycoumarin-4-acetyl (Mca) is released and its fluorescence can be used to quantify MMP activity. Mca displays excitation/emission maxima of 328/420 nm, respectively.
Z-(L-Arg)-AMC is a fluorogenic substrate for trypsin, cathepsin B, and cathepsin H.1,2Upon enzymatic cleavage by trypsin, cathepsin B, or cathepsin H, 7-amino-4-methylcoumarin (AMC) is released and its fluorescence can be used to quantify trypsin, cathepsin B, and cathepsin H activity. AMC displays excitation/emission maxima of 340-360/440-460 nm, respectively.
1.Zimmerman, M., Ashe, B., Yurewicz, E.C., et al.Sensitive assays for trypsin, elastase, and chymotrypsin using new fluorogenic substratesAnal. Biochem.78(1)47-51(1977) 2.Brindley, P.J., Kalinna, B.H., Dalton, J.P., et al.Proteolytic degradation of host hemoglobin by schistosomesMol. Biochem. Parasitol.89(1)1-9(1997)
L-Leu-AMC is a fluorogenic substrate for leucine aminopeptidase.1Upon enzymatic cleavage by leucine aminopeptidase, AMC is released and its fluorescence can be used to quantify leucine aminopeptidase activity. AMC displays excitation/emission maxima of 340-360/440-460 nm, respectively.
1.Izquierdo, M., Lin, D., O’Neill, S., et al.Development of a high-throughput screening assay to identify inhibitors of the major M17-leucyl aminopeptidase from Trypanosoma cruzi using rapidfire mass spectrometrySLAS Discov.25(9)1064-1071(2020)
Ac-ANW-AMC is a fluorogenic substrate for the β5i/LMP7 subunit of the 20S immunoproteasome. [1] Upon cleavage, 7-amino-4-methylcoumarin (AMC) is released and its fluorescence can be used to quantify the activity of the β5i/LMP7 subunit of the 20S immunoproteasome. AMC displays excitation/emission maxima of 340-360/440-460 nm, respectively.
Reference[1]. Winter, M.B., La Greca, F., Arastu-Kapur, S., et al. Immunoproteasome functions explained by divergence in cleavage specificity and regulation. eLife 6:e27364, (2017).
4-Methylumbelliferyl β-D-Galactopyranoside-6-sulfate (sodium salt) (4-MU-Gal-6S) is a fluorogenic substrate used to quantify N-acetylgalactosamine-6-sulphatase (GALNS) activity. 4-MU-Gal-6S is cleaved by GALNS to release the fluorescent moiety 4-MU. 4-MU fluorescence is pH-dependent with excitation maxima of 320 and 360 nm at low (1.97-6.72) and high (7.12-10.3) pH, respectively, and an emission maximum ranging from 445 to 455 nM, increasing as pH decreases. It has been used to detect Morquio disease type A, a lysosomal storage disorder in which GALNS is deficient. 4-MU-Gal-6S can be used to assess GALNS activity in a very small blood volume to determine the extent of deficiency.
4-Methylumbelliferyl 2-sulfamino-2-deoxy-α-D-Glucopyranoside (4-MU-α-GlcNS) is a fluorogenic substrate of heparin sulphamidase. Heparin sulphamidase cleaves 4-MU-α-GlcNS to yield 4-MU-α-GlcNH2, which is then cleaved by α-glucosaminidase to release the fluorescent product 4-MU, which displays an emission maxima of 445-454 nm. The excitation maxima for 4-MU is pH-dependent: 330, 370, and 385 nm at pH 4.6, 7.4, and 10.4, respectively. 4-MU-α-GlcNS has been used to quantify heparin sulphamidase deficiencies associated with Mucopolisaccaridosis IIIA and other lysosomal disorders.
4-Methylumbelliferyl-β-D-galactoside (MUG) is a fluorogenic substrate for β-galactosidase (β-gal).1Hydrolysis of MUG by β-gal releases the fluorescent moiety 4-MU, which displays a pH-dependent excitation maximum of 330, 370, and 385 nm at pH 4.6, 7.4, and 10.4, respectively, and an emission maximum between 445-454 nm.2MUG has been used to detect β-gal activity in intact bacteria, yeast, and mammalian cells.1
1.Vidal-Aroca, F., Giannattasio, M., Brunelli, E., et al.One-step high-throughput assay for quantitative detection of β-galactosidase activity in intact Gram-negative bacteria, yeast, and mammalian cellsBiotechniques40(4)433-434(2006) 2.Zhi, H., Wang, J., Wang, S., et al.Fluorescent properties of hymecromone and fluorimetric analysis of hymecromone in compound dantong capsuleJ. Spectrosc.1(1)147128(2013)
Mca-Ala-Pro-Lys(Dnp)-OH, a specific quenched fluorogenic substrate for ACE2, enables the detection of ACE2 activity in various tissues including urinary, heart, and lung.
Coumberone is a metabolic fluorogenic probe and isoform-selective substrate for all AKR1C isoforms. It can be reduced by all four members of the AKR1C family to its fluorescent alcohol coumberol. Coumberone is a valuable tool for researching AKR1C.