Fluorogenic Substrates

Acetyl-L-alanine 7-amido-4-methylcoumarin is a fluorogenic substrate for aminopeptidase. The enzyme cleaves the N-terminal amino acids, yielding a blue fluorescent solution.

Fluorogenic substrate for ?-arabinofuranosidase

4-Methylumbelliferyl palmitate, commonly names as 4-MUP, is a fluorescent probe that binds to lipases, enzymes responsible to assist on the hydrolysis of fats. In diagnosis, 4-MUP is used for screening of lysosomal storage diseases (LSD) caused by a deficiency of a single enzyme involved in the metabolism of glycoproteins, lipids or mucopolysaccharides.

A sensitive fluorogenic substrate for fluorometric and UV-spectrophotometric assays of phosphatases. It is used for the detection of C. perfringens because acid phosphatase is a highly specific indicator for C. perfringens.

Glutaryl-glycyl-L-arginine 7-amido-4-methylcoumarin hydrochloride is a synthetic peptide substrate used as a fluorogenic substrate for aminopeptidase.

Fluorogenic substrate for beta-glucuronidase GUS

4-Methylumbelliferyl 2-acetamido-2-deoxy-beta-D-glucopyranoside-6-sulfate potassium salt is a fluorogenic substrate for beta-N-acetylglucosaminidase. After enzymatic cleaveage, free 4-methylumbelliferone (also known as hymecromone) is released, exhibiting blue fluorescence upon excitation with UV light. The strongest fluorescence of 4-methylumbelliferone requires deprotonation of the hydroxyl group (thus requires alkaline pH), with a maximal fluorescence intensity obtained with excitation at 350 to 370 nm and emission at 440 to 470 nm. The use of 4-methylumbelliferyl 2-acetamido-2-deoxy-beta-D-glucopyranoside-6-sulfate potassium saltas a substrate for measuring the beta-N-acetylglucosaminidase activity is used for diagnosis of Mucopolysaccharidosis IIIB (MPS IIIB) (a rare lysosomal storage disorder) and Sanfilippo B syndrome screening.

N-alpha-CBZ-L-Arginine 7-amido-4-methylcoumarin hydrochloride is a Ffluorogenic substrate used in protease activity assays.

4-Methylumbelliferyl-alpha-L-iduronic acid, cyclohexylammonium salt is a fluorogenic substrate for the detection of alpha-L-iduronidase activity. It has been used to measure the activity of this enzyme in cells and tissues and as an indicator of tissue damage. The product is also used in food testing, diagnostics, and environmental testing.

The DDAO molecule [7-hydroxy-9H-(1, 3-dichloro-9, 9 dimethylacridin-2-one] is used as a stain and a dye in molecular biology, due to its ability to bind intracellular and extracellular DNA. DDAO galactoside is a fluorogenic substrate for galactosidase.

L-Serine 7-amido-4-methylcoumarin hydrochloride is a fluorogenic peptide substrate for peptidases. This AMC peptide substrate is normally used to rapidly profile the N-terminal specificity of proteases.

6-Hexadecanoylamino-4-methylumbelliferyl phosphorylcholine (HMUPC) is a fluorogenic substrate of acid sphingomyelinase (ASM) in the application of the diagnosis of Niemann–Pick type A and B (NPAB). NPAB is one of the lysosomal storage diseases, caused by a deficiency of ASM. This deficiency results in the accumulation of sphingomyelin and other lipids in various tissues. The assay using HMUPC as the substrate shows rapid and robust determination of ASM activity. When combined with the unlabelled natural substrate such as sphingomyelin as a specific inhibitor of ASM activity, the specificity of assay is enhanced up to the level of the classical assay with radiolabelled natural substrate. HMUPC is also used as the substrate in the ASM assay to investigate the retention of lysosomal enzyme activity of bi-functional IgG-ASM fusion protein in the delivery across blood-brain barrier.