fluorogenic

Enteropeptidase fluorogenic substrate is a substrate for enteropeptidase that contains a 7-amino-4-trifluoromethylcoumarin (AFC) moiety. Enteropeptidase is a serine protease expressed in the proximal small intestine of higher animals that converts inactive trypsinogen to active trypsin by endoproteolytic cleavage. Enteropeptidase recognizes the highly specific amino acid sequence DDDDK on the fluorogenic substrate and cleaves after the lysine residue, releasing the AFC moiety. Enteropeptidase activity is quantified by fluorescent detection of AFC, which displays excitation/emission spectra of 380/500 nm.

Enteropeptidase fluorogenic substrate is a substrate for enteropeptidase that contains a 7-amino-4-trifluoromethylcoumarin (AFC) moiety. Enteropeptidase is a serine protease expressed in the proximal small intestine of higher animals that converts inactive trypsinogen to active trypsin by endoproteolytic cleavage.1,2Enteropeptidase recognizes the highly specific amino acid sequence DDDDK on the fluorogenic substrate and cleaves after the lysine residue, releasing the AFC moiety. Enteropeptidase activity is quantified by fluorescent detection of AFC, which displays excitation/emission spectra of 380/500 nm.3

The renin fluorogenic substrate consists of the normal peptide substrate for renin which has been linked to the fluorophore EDANS at one end and to a non-fluorescent quenching molecule (Dabcyl) at the other. After cleavage by renin, the product (peptide-EDANS) is brightly fluorescent and can be easily analyzed using an excitation wavelength of 340 nm and emission wavelengths of 485-510 nm.

4-Methylumbelliferyl-β-D-Glucopyranoside (4-MU-GLU) is used in the GCase activity assay based on the catalytic hydrolysis of 4-methylumbelliferyl β-D-glucopyranoside that releases the highly fluorescent 4-methylumbelliferyl (4-MU).

Maleimide (2,5-Pyrroledione) (2,5-Pyrroledone) is a new nanoparticle surface functional group which favors easy conjugation with cell penetration peptides. The conjugation is enabled via click chemistry to preserve its biofunctions.

Cathepsin D and E FRET Substrate acetate is a fluorogenic substrate for cathepsins D and E but not for B, H, or L. Cathepsin D and E FRET Substrate acetate can be used in mechanistic studies on cathepsins D and E studies.

Z-Gly-Gly-Arg-AMC TFA (66216-78-2 free base) is a thrombin-specific fluorogenic substrate used to assess thrombin generation in platelet-rich plasma (PRP) and platelet-poor plasma (PPP).

ERAP1-IN-1 is an endoplasmic reticulum aminopeptidase 1 (ERAP1) inhibitor that allosterically activates ERAP1’s hydrolysis of fluorogenic and chromogenic amino acid substrates, while competitively inhibiting its activity toward a nonamer peptide representative of physiological substrates.

H-β-Ala-AMC TFA is a substrate for aminopeptidase and serves as a building block for synthesizing the fluorogenic vanin I substrate [pantothenate-AMC].

4-Methylumbelliferyl-α-D-Galactopyranoside (4MU-α-Gal) (4-MU-α-Gal) is a fluorogenic substrate of α-galactosidase. In addition to its use in characterizing novel α-galactosidases, 4-MU-α-Gal is used in assays to evaluate deficiency in α-galactosidase activity related to Fabry disease.

Ac-DEVD-AFC, a fluorogenic substrate, undergoes cleavage by caspase-3, producing a blue fluorescent compound upon excitation at 400 nm, which can be quantified at an emission maximum of 505 nm.

BOP-JF549 is a fluorescent dual α9β1/α4β1 integrin inhibitor. Comprises BOP conjugated to Janelia Fluor® 549. Fluorogenic: fluoresces only once bound to integrins, enabling hassle-free no-wash experiments. Bright and photostable, enabling live cell tracking of integrin receptors over long time-course experiments. Excitation maximum = 549 nm; emission maximum = 571 nm.

Dnp-PLGMWSR is a fluorogenic substrate for matrix metalloproteinase-2 (MMP-2) and MMP-9. The activity of MMP-2 and MMP-9 can be quantified by measuring tryptophan fluorescence that is unquenched upon peptide hydrolysis that removes the N-terminal dinitrophenol (Dnp) group.

7-Lysylalanyl-4-methylcoumarinamide is a Fluorogenic substrate for dipeptidyl aminopeptidase.

Dnp-PLALWAR is a fluorogenic substrate for matrix metalloproteinase-1 (MMP-1) and MMP-8. The activity of MMP-1 and MMP-8 can be quantified by measuring tryptophan fluorescence that is unquenched upon peptide hydrolysis that removes the N-terminal dinitrophenol (Dnp) group.

CrBKA, a fluorogenic small-molecule substrate of SIRT6, exhibits weak activity [1].

Boc-Leu-Gly-Arg-AMC is a fluorogenic [AMC] substrate employed in enzymatic assays to study convertases.

HIV Protease Substrate 1 TFA, a fluorogenic substrate for HIV protease, facilitates the investigation of the enzyme's activity [1].

Ac-WEHD-AFC TFA is a fluorogenic substrate for caspase-1, used to measure the enzyme's activity in tumor and inflammation research [1].

TNO211 is a bioactive peptide that functions as a highly soluble fluorogenic substrate for select Matrix Metalloproteinases (MMPs)—specifically MMP-2, 8, 12, 13, and 14. Incorporating a cleavable Gly-Leu bond and an EDANS/DABCYL fluorophore/quencher pair, TNO211 enables sensitive detection of MMP activity through fluorescence assays, with an absorption/emission profile at 340/490 nm. It is particularly effective in analyzing MMPs in culture media from endothelial cells as well as untreated synovial fluid from patients, highlighting its utility in investigations of MMP-associated cellular behaviors, including proliferation, migration (adhesion/dispersion), differentiation, angiogenesis, apoptosis, and host defense.

4-Methylumbelliferyl caprylate (MUCAP) is a fluorogenic substrate for C8 esterase. MUCAP is cleaved by C8 esterase to release the fluorescent moiety 4-methylumbelliferyl (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.

Ac-Leu-Arg-AMC (Acetyl-Leucyl-Arginyl-7-amido-4-methylcoumarin) is a fluorogenic peptide substrate.

Dabcyl-KTSAVLQSGFRKME-Edans TFA, a fluorogenic peptide, serves as a substrate for quantifying protease enzymatic activities. This compound is also under investigation for its applicability in studying 2019-nCoV (COVID-19) infection [1] [2] [3].

WH-15 is a fluorogenic phospholipase C (PLC) reporter, exhibiting Michaelis-Menten constant (Km) values of 49, 30, and 86.1 µM for PLC-γ1, PLC-δ1, and PLC-β2, respectively. It undergoes enzymatic cleavage in a cascade reaction to produce fluorescent 6-aminoquinoline. This compound enables the imaging of PLC activity in live cells [1].

4-Methylumbelliferyl β-D-N,N'-diacetylchitobioside (4-μU-(GlcNAc)2) is a fluorogenic substrate for chitinases and chitobiosidases. 4-μU-(GlcNAc)2 is cleaved by chitinases and chitobiosidases to release the fluorescent moiety 4-μU. 4-μU fluorescence is pH-dependent with excitation maxima of 320 and 360 nm at low (1.97-6.72) and high pH (7.12-10.3), respectively, and an emission maximum ranging from 445 to 455 nm, increasing as pH decreases.

Suc-YVAD-AMC is a fluorogenic substrate for caspase-1. Upon enzymatic cleavage by caspase-1, 7-amino-4-methylcoumarin (AMC) is released and its fluorescence can be used to quantify caspase-1 activity. AMC displays excitation/emission maxima of 340-360/440-460 nm, respectively.

M-2420 is a fluorogenic substrate specifically designed for the β-secretase site associated with the Swedish mutation of the amyloid precursor protein (APP).

7-Acetoxy-1-methylquinolinium iodide (AMQI) serves as a fluorogenic substrate for cholinesterase, with excitation (Ex) at 320 nm and emission (Em) at 410 nm. In a fluorometric flow system, the hydrolysis of AMQI is used to detect and identify inhibitors.

Cathepsin D and E FRET Substrate acetate is a fluorogenic substrate selectively cleaved by cathepsins D and E at the Phe-Phe amide bond, demonstrating no activity towards cathepsins B, H, or L. This specificity renders it essential for routine assays and mechanistic studies focusing on cathepsins E and D[1].

BOP-JF646 is a fluorescent dual α9β1/α4β1 integrin inhibitor. Comprises BOP conjugated to Janelia Fluor®646. Fluorogenic: fluoresces only once bound to integrins, enabling hassle-free no-wash experiments. Bright and photostable, enabling live cell tracking of integrin receptors over long time-course experiments. Excitation maximum = 655 nm; emission maximum = 672 nm.

Boc-LRR-AMC is a fluorogenic substrate for the trypsin-like activity of the 26S proteasome or 20S proteolytic core. Upon enzymatic cleavage by the 26S proteasome or 20S proteolytic core, amino-4-methylcoumarin (AMC) is released and its fluorescence can be used to quantify 26S proteasome or 20S proteolytic core trypsin-like activity. AMC displays excitation/emission maxima of 340-360/440-460 nm, respectively.

4-Methylumbelliferyl-α-L-iduronide (free acid) is a fluorogenic substrate for α-L-iduronidase, an enzyme found in cell lysosomes that is involved in the degradation of glycosaminoglycans such as dermatan sulfate and heparin sulfate. 4-Methylumbelliferyl-α-L-iduronide is cleaved by α-L-iduronidase to release the fluorescent moiety 4-methylumbelliferyl (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. This substrate is used in assays that measure the activity of α-L-iduronidase, which is commonly deficient in a type of lysosomal storage disease called mucopolysaccharidosis.

HX103, an epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor (TKI)-based fluorogenic probe, exhibits selective inhibition towards various EGFR mutations, including EGFR 19 del, EGFR L858R, EGFR wild type, and EGFR T790M, with respective IC50 values of 1.3, 1.5, 4.0, and 977 nM. It is utilized for quantifying active-EGFR to foresee agent sensitivity in NSCLC (non-small cell lung cancer) patients harboring EGFR-activating mutations [1].

Dnp-PLAYWAR is a fluorogenic substrate for matrix metalloproteinase-8 (MMP-8) and MMP-26. The activity of MMP-8 and MMP-26 can be quantified by measuring tryptophan fluorescence that is unquenched upon peptide hydrolysis that removes the N-terminal dinitrophenol (Dnp) group.

Z-VAD-AMC is a fluorogenic substrate for caspase-1. Upon enzymatic cleavage by caspase-1, 7-amino-4-methylcoumarin (AMC) is released and its fluorescence can be used to quantify caspase-1 activity. AMC displays excitation/emission maxima of 340-360/440-460 nm, respectively.

Ac-VEID-AMC is a fluorogenic substrate based on the caspase-6 cleavage site in lamin A at amino acids VEID during apoptosis.1It has also been reported to be cleaved by related proteases, including caspase-8.2Caspase activity can be quantified by fluorescent detection of free AMC (also known as 7-amino-4-methylcoumarin), which is excited at 340-360 nm and emits at 440-460 nm. 1.Talanian, R.V., Quinlan, C., Trautz, S., et al.Substrate specificities of caspase family proteasesJ. Biol. Chem.272(15)9677-9682(1997) 2.Chae, H.J., Park, K.M., Lee, G.Y., et al.Je-Chun-Jun induced apoptosis of human cervical carcinoma HeLa cellsActa Pharmacologica Sinica25(10)1372-1379(2004)

Dnp-RPLALWRS is a fluorogenic substrate for matrix metalloproteinase-7 (MMP-7). The activity of MMP-7 can be quantified by measuring tryptophan fluorescence that is unquenched upon peptide hydrolysis that removes the N-terminal dinitrophenol (Dnp) group.

Dnp-P-Cha-G-Cys(Me)-HA-K(Nma)-NH2 is a fluorogenic substrate for matrix metalloproteinase-1 (MMP-1) and MMP-9. Upon cleavage by MMP-1 or MMP-9, N-methylanthranilic acid (Nma) is unquenched and its fluorescence can be used to quantify MMP activity. Nma displays excitation/emission spectra of 340/440 nm, respectively.

HIV Protease Substrate 1 is a fluorogenic compound that offers insights into the enzymatic activity of HIV protease, making it a valuable tool for studying this aspect.

Ac-PAL-AMC is a fluorogenic substrate for the β1i LMP2 subunit of the 20S immunoproteasome. Upon cleavage, 7-amino-4-methylcoumarin (AMC) is released, and its fluorescence can quantify the activity of the β1i LMP2 subunit. Ac-PAL-AMC is selective for the immunoproteasome over the constitutive proteasome. AMC displays excitation emission maxima of 351 430 nm.

4-Methylumbelliferyl-2-acetamido-2-deoxy-β-D-glucopyranoside is a fluorogenic substrate for β-hexosaminidases. Upon enzymatic cleavage by β-hexosaminidases, 4-methylumbelliferone (4-μU) is released, and its fluorescence quantifies β-hexosaminidase activity. 4-MU fluorescence is pH-dependent with excitation maxima of 320 and 360 nm at low (1.97-6.72) and high pH (7.12-10.3), respectively, and an emission maximum ranging from 445 to 455 nm, increasing as pH decreases. This compound has been used to quantify β-hexosaminidase activity in serum or leukocytes from patients with GM2 gangliosidoses, such as Tay-Sachs disease, which is characterized by defects in the α subunit of β-hexosaminidase.

Ac-RGK(Ac)-AMC, fluorogenic substrate for assaying histone deacetylase (HDAC) activity in a two-step enzymatic reaction. The assay consists of the initial lysine deacetylation by HDAC followed by the release of the fluorescent group by trypsin.

Z-Gly-Gly-Arg-AMC is a specific fluorogenic substrate used to assess thrombin generation and thrombin activity in Platelet-Rich Plasma (PRP) and Platelet-Poor Plasma (PPP).

Mca-VDQVDGW-Lys(Dnp)-NH2, a fluorogenic substrate specific to caspase-7, facilitates the quantification of caspase-7 activity.

4-Methylumbelliferyl-β-D-N,N',N''-triacetylchitotrioside is a fluorogenic substrate for chitinases and chitotriosidases. 4-Methylumbelliferyl-β-D-N,N',N''-triacetylchitotrioside is cleaved by chitinases and chitotriosidases to release the fluorescent moiety 4-methylumbelliferyl (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 screen plasma samples for reduced chitotriosidase activity that may be indicative of lysosomal storage disorders.

Boc-Arg-Val-Arg-Arg-AMC hydrochloride (Boc-RVRR-AMC) is a synthetic fluorogenic substrate efficiently cleaved by the enzyme furin.

Phe-Pro-Arg-PABA-Resorufin is a Chromogenic and fluorogenic peptide substrate for the highly sensitive detection of proteases in biological matrices. The substrate is also applicable to the sensitive detection of the thrombin inhibitor dabigatran in human

Mu-6S-Palm-β-Glc is a fluorogenic substrate of palmitoyl-protein thioesterase (PPT, also known as CLN1), a lysosomal hydrolase that removes long-chain fatty acyl groups from modified cysteine residues in proteins. Upon cleavage by PPT CLN1, it releases the fluorescent moiety 4-methylumbelliferyl (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. This substrate is used in assays measuring PPT activity, commonly deficient in the neurodegenerative disorder known as infantile neuronal ceroid lipofuscinosis.

Boc-Ile-Glu-Gly-Arg-AMC (IEGR-AMC) is a specific, highly fluorogenic substrate for clotting enzyme factor Xa (coagulation factor Xa) and is also hydrolyzed by acrosin from the ascidian Halocynthia roretzi.

Gly-Arg-AMC is a fluorogenic substrate for cathepsin C.1 Upon enzymatic cleavage by cathepsin C, 7-amino-4-methylcoumarin (AMC) is released and its fluorescence can be used to quantify cathepsin C activity. AMC displays excitation/emission maxima of 340-360/440-460 nm, respectively. |1. Rubach, J.K., Cui, G., Schneck, J.L., et al. The amino-acid substituents of dipeptide substrates of cathepsin C can determine the rate-limiting steps of catalysis. Biochemistry 51(38), 7551-7568 (2012).