Fluorogenic Substrates

Resorufin b-D-glucopyranoside is a stable red fluorogenic galactosidase substrate.

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.

4-Methylumbelliferyl lignocerate is a fluorogenic substrate for lipase. 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 lignocerateas a substrate for measuring the lipase activity is used for X-linked adrenoleukodystrophy diagnosis and peroxisomal disorder screening.

Living cells of Escherichia coli can be reliably detected with media that contain a chromogenic or fluorogenic substrate for beta-glucuronidase (also termed β-D-glucuronidase), an enzyme that occurs almost exclusively in E. coli. RUG®, Resorufin-beta-D-glucuronic acid methyl ester is a highly sensitive chromogenic and fluorogenic indicator for E. coli. After removal of the methyl ester group by unspecific hydrolysis, orange colored product is cleaved by beta-glucuronidase in liquid media or on agar plates and yields an intense pink color. Red fluorescence is obtained upon excitation at 560-570 nm.Formally ER175455.

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.

A substrate used for the diagnosis of Sanfilippo disease C (MPS III C).

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.

4-Methylumbelliferyl-beta-D-glucuronic acid methyl ester is a chromogenic substrate for β-glucuronidase. It is used in diagnostic applications and as a substrate in enzyme kinetics experiments. 4-Methylumbelliferyl-beta-D-glucuronic acid methyl ester has fluorescence properties that allow it to be detected by chemiluminescence and bioluminescence.

Resorufin-7-O-phosphate is a fluorogenic substrate for alkaline phosphatase that exhibits a red color after cleavage that can be measured at ~ 585nm.

Fluorescent dye substrate for flow cytometric measurements