fluorogenic substrate

EDANS (EDANS acid [5-((2-Aminoethyl)amino)naphthalene-1-sulfonic acid]) is a novel and quenched fluorogenic substrate for assaying retroviral protease by resonance energy transfer (RET).

hCYP3A4 Fluorogenic substrate 1, a phthalimide derivative, exhibits low activity against COX.

MMP-3 Fluorogenic Substrate, a compound designed for matrix metalloproteinase-3 (MMP-3) activity quantification, releases 7-methoxycoumarin-4-acetyl (Mca) upon MMP-3 cleavage. Mca's fluorescence, with excitation and emission maxima at 328 and 420 nm respectively, serves as a measurable indicator of MMP-3 activity.

SGPL1 fluorogenic substrate is a fluorogenic substrate of sphingosine-1-phosphate lyase (SGPL1) used to measure SGPL1 activity [1].

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).

4-Methylumbelliferyl-β-D-glucuronide hydrate (MUG) is a fluorogenic substrate (λex=362 nm, λem=445 nm).

4-Methylumbelliferyl phosphate (4-MUP) (4-MUP) is used as a fluorogenic substrate of alkaline phosphatases.

ADHP (10-Acetyl-3,7-dihydroxyphenoxazine) is a fluorogenic peroxidase substrate (λex=530 nm, λem=590 nm).

CUG is a fluorogenic substrate (λex=386, λem=445 nm, ε=32K).

Fluorescein di(β-D-galactopyranoside) is a fluorogenic substrate for β-galactosidase (λem=535 nm, λex=485 nm).

PL553 is a specific and high-affinity fluorogenic substrate of Leukotriene A4 hydrolase [λmax: 210 nm; λem: 410 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 is a styrene-conjugated fluorogenic probe for radical-trapping antioxidant (RTA) activity.1 Co-autoxidation of the STY-BODIPY signal carrier and a hydrocarbon co-substrate can be quantified by monitoring the loss of absorbance at 571 nm. STY-BODIPY has been used to measure the activity of RTAs, as well as the kinetics and stoichiometry of RTA reactions in cell-free assays.1,2,3References1. Haidasz, E.A., Van Kessel, A.T.M., and Pratt, D.A. A continuous visible light spectrophotometric approach to accurately determine the reactivity of radical-trapping antioxidants. J. Org. Chem. 81(3), 737-744 (2016).2. Shah, R., Shchepinov, M.S., and Pratt, D.A. Resolving the role of lipoxygenases in the initiation and execution of ferroptosis. ACS Cent. Sci. 4(3), 387-396 (2018).3. Chauvin, J.-P.R., Haidasz, E.A., Griesser, M., et al. Polysulfide-1-oxides react with peroxyl radicals as quickly as hindered phenolic antioxidants and do so by a surprising concerted homolytic substitution. Chem. Sci. 7(10), 6347-6356 (2016). STY-BODIPY is a styrene-conjugated fluorogenic probe for radical-trapping antioxidant (RTA) activity.1 Co-autoxidation of the STY-BODIPY signal carrier and a hydrocarbon co-substrate can be quantified by monitoring the loss of absorbance at 571 nm. STY-BODIPY has been used to measure the activity of RTAs, as well as the kinetics and stoichiometry of RTA reactions in cell-free assays.1,2,3 References1. Haidasz, E.A., Van Kessel, A.T.M., and Pratt, D.A. A continuous visible light spectrophotometric approach to accurately determine the reactivity of radical-trapping antioxidants. J. Org. Chem. 81(3), 737-744 (2016).2. Shah, R., Shchepinov, M.S., and Pratt, D.A. Resolving the role of lipoxygenases in the initiation and execution of ferroptosis. ACS Cent. Sci. 4(3), 387-396 (2018).3. Chauvin, J.-P.R., Haidasz, E.A., Griesser, M., et al. Polysulfide-1-oxides react with peroxyl radicals as quickly as hindered phenolic antioxidants and do so by a surprising concerted homolytic substitution. Chem. Sci. 7(10), 6347-6356 (2016).

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 quantifies the activity of the β5i LMP7 subunit. AMC displays excitation emission maxima of 340-360 440-460 nm, respectively.

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.

4-Methylumbelliferyl phosphate disodium (4-MUP), an anionic organophosphate, serves as a fluorogenic substrate for acid and alkaline phosphatases. Additionally, this compound acts as a simulant for nerve agents [1] [2] [3].

DABCYL-Glu-Arg-Nle-Phe-Leu-Ser-Phe-Pro-EDANS is a fluorescent dye used as a fluorogenic substrate for aspartyl proteinase from the human malaria parasite [1]. It should be stored protected from light.