Enzyme Substrates

2-Naphthyl beta-D-galactopyranoside is a glycoside that is found in the flowers of the plant Eschscholzia californica. It is used as a localizer for bacterial strains and can be hydrolyzed by enzymes to release naphthalene. 2-Naphthyl beta-D-galactopyranoside has been shown to inhibit the growth of corynebacterium and diazonium salt, which may be due to its ability to bind to DNA and disrupt protein synthesis. The potential use of this molecule is for its insolubility, which prevents it from being absorbed by skin cells.

Stability Hygroscopic Applications Labelled analogue of γ-Butyrobetaine, a synthetic carnitine related compound used as a transporter substrate in the cloning and sequencing of human carnitine transporter 2 (CT2). References Enomoto, A. et al.: J. Biol. Chem., 277, 36262 (2002);

5-bromo-3-indolyl-b-d-glucopyranoside, also known as 5BIG, is an enzyme substrate for beta-glucosidase. It is used in bacterial assays for the identifcation of Gram-positive bacteria such as Staphylococcus aureus and Bacillus subtilis. After cleavege, it gives an intense blue color.

3-HTC reacts reversibly with thiols and disulfides, and can be used to measure dynamic GSH/GSSH ratios in vitro as well as to monitor the reversible redox status of whole cell lysates.(λmax: 448 nm in its reduced thiolate form, and a λmax 370-410 nm for the oxidized mixed disulfide).

5-Bromo-3-indolyl 2-acetamido-2-deoxy-b-D-glucopyranoside is a lapis substrate, a chromogenic substrate that yields a deep blue color after cleavage. It is used in lysosomal storage disorder screening such as for Sanfilippo syndrome.

2-Nitrophenyl-1-thio-beta-D-galactopyranoside is a nitroaryl compound that can be synthesized from beta-D-galactopyranosides and nitric acid. It has a molecular conformation that is stabilized by hydrogen bonds, which allows it to form supramolecular assemblies with other molecules. The nitro group in this molecule is responsible for the formation of the nitro group in 1,2,4,5,-tetranitrobenzene.

4-Nitrophenyl 2-acetamido-3-O-(3,4,6-tri-O-acetyl-2-deoxy-2-phthalimido-b-D-glucopyranosyl)-2-deoxy -a -D -galactopyranoside is a Fluorogenic substrate that is used in the detection of beta glucuronidase activity. It can be used for food testing, staining, environmental testing and chromogenic substrate for diagnostic purposes. It has a CAS number of 184840-55-9 and is available in high purity and quality. 4NPPAG is an enzyme substrate that is used to measure the amount of beta glucuronidase activity in culture media by chemiluminescence or bioluminescence.

Fluorogenic substrate targeting esterase and lipase

4-Methylumbelliferyl alpha-L-idopyranosiduronic acid 2-sulfate disodium salt is a chromogenic substrate that is used in the detection of glycosidases. It reacts with enzymes such as alpha-amylase, beta-galactosidase, and beta-glucosidase to produce a fluorescent product. This product can be detected using a fluorometer or spectrophotometer. The CAS number for 4-methylumbelliferyl alpha-L-idopyranosiduronic acid 2-sulfate disodium salt is 1045020-74-3.

DMNPE-caged Luciferin is a photoactivatable probe that can easily penetrate the cell membrane, and by exposure to UV light it undergoes a photochemical decomposition liberating a Luciferin molecule. Once exposed to Luciferase enzyme expressed by the cells, Luciferin emits visible (and measurable) light. Using_x000D_this technique, imaging of luciferase activity was performed both in living cells and in mouse tumor xenograft, by intraperitoneal injection of the caged DMNPE luciferin (Chem. Commun., 2009, 0, 4028-4030).

2-Chloro-4-nitrophenyl β-D-cellotrioside is a chromogenic substrate use in cellulase activity assays. This enzyme, which is responsible to break down cellulose, hydrolyzes 2-chloro-4-nitrophenyl beta-D-cellotrioside and releases 2-chloro-4-nitrophenol than can be quantified spectrophotometrically. The use of 2-chloro-4-nitrophenyl beta-D-cellotrioside as substrate for enzymatic assays allows detecting bacterial cellulose in urine tract infections in the biofilm state.

3,3'-Diaminobenzidine tetrahydrochloride or DAB hydrochloride is the hydrochloride salt of biphenyl tetra-aniline. DAB hydrochloride is a water-soluble formulation of DAB and is used in immunohistochemical staining of nucleic acids and proteins. DAB hydrochloride is used in Alzheimer's disease research, Aβ protein amyloid plaques are targeted by a primary antibody, and subsequently by a secondary antibody, which is conjugated with a peroxidase enzyme. This will bind DAB as a substrate and oxidize causing a colour change. 3,3’-Diaminobenzidine tetrahydrochloride is also available as the tetrahydrochloride hydrate and as the free base.

L-Luciferin is an enzyme substrate that is used in the detection of bioluminescence. It is a fluorogenic substrate that can be used to detect the presence of glucose in urine. L-Luciferin is also a chemiluminescent substrate that can be used to detect ATP, which is required for cell metabolism. L-Luciferin has been shown to be stable at pH levels ranging from 2.5 to 9 and can be stored for up to 3 years when stored at 4°C with no light or oxygen exposure. This product has CAS No. 754978-59-1, indicating high purity and quality. L-Luciferin will not stain or react with other compounds, making it highly suitable for use as a conjugate in diagnostic tests or as a ligand in fluorescent staining reactions.

5-Bromo-4-chloro-3-indoxyl-alpha-D-N-acetylneuraminic acid, cyclohexylammonium salt is a chemiluminescent substrate that provides a significant increase in light emission. It has a CAS number of 59820-92-0 and is supplied in high purity and high quality. This product is used for the detection of various biological reactions, including diagnostics, enzyme substrates, food testing, and more.

4MU-alpha-L-idopyranosiduronic acid is a fluorogenic substrate for alpha-L-iduronidase. Active alpha-L-iduronidase cleaves the MU moiety from the synthetic substrate, which results in release of a fluorescent signal (Ex 360 nm / Em 455 nm). The reaction is stopped by the addition of sodium carbonate. 4MU-alpha-L-idopyranosiduronic acid is used in alpha-L-iduronidase assays in the context of biochemical characterisation of samples from patients affected by lysosomal storage disease mucopolysaccharidosis I (MPS I).

Butyl 4-nitrophenyl hexyl phosphonate is a lipase suicide inhibitor that is a chromogenic substrate for lipases which upon enzymatic action, binds irreversibly to the lipase and inactivates it. The substrate can be used for the determination of active sites in lipases, especially in lipases preferring long chain leaving groups.

Chromogenic substrate for α-amylase yielding a yellow solution upon cleavage.

Chromogenic substrate for N-acetylgalactosaminase

Aldol 491 acetate is a food and environmental testing chemical, which can be used for the detection of bacteria. Aldol 491 acetate provides a chromogenic substrate for the detection of enzymes and can be used as a substrate in enzyme-linked immunosorbent assay (ELISA) procedures. Aldol 491 acetate also has chemiluminescent properties and can be used as a fluorogenic substrate.

The 4-Methylumbelliferyl b-D-ribofuranoside is a top-notch fluorogenic substrate for glycosidase activity detection. Upon hydrolysis by enzymes, it generates a strongly fluorescent 4-methylumbelliferone molecule, enabling rapid assay results and real-time monitoring of enzymatic reactions. This product is suitable for many biological research applications, such as in vitro analysis, diagnostic testing, and drug discovery. With its high sensitivity and specificity, it delivers consistent and accurate results, making it an essential tool for scientists and researchers.

3,3'-Diaminobenzidine (DAB) is an organic compound derived from benzidine. DAB is water-soluble as the tetrahydrochloride salt and is used in immunohistochemical staining of nucleic acids and proteins. Within research in Alzheimer's disease, Aβ protein amyloid plaques are targeted by a primary antibody, and subsequently by a secondary antibody, which is conjugated with a peroxidase enzyme. This will bind DAB as a substrate and oxidize it, producing an easily observable brown colour. 3,3’-Diaminobenzidine tetrahydrochloride hydrate is also available as the tetrahydrochloride salt and as the free base.

As an alternative to commonly used colorimetric and fluorescent substrates, 3-(2'-Spiroadamantane)-4-methoxy-4-(3''-phosphoryloxy)phenyl-1,2-dioxetane, abbreviated AMPPD, is a chemiluminescent substrate for alkaline phosphatase (ALP). It allows detection of alkaline phosphatase at a high sensitivity. Literature reports that the co-presence of 0.1% bovine serum albumin (BSA) leads to a signal increase of 10-folds under selected conditions.

5-Fluorescein isothiocyanate isomer I (5FITC) is a fluorescent probe that has been used to study enzyme activities in human serum. 5FITC binds to the active site of many enzymes, including dopamine β-hydroxylase, and can be detected by measuring the fluorescence intensity. The optimum concentration for this enzyme form is 10 nM for dopamine β-hydroxylase and 20 nM for brain functions. Mouse monoclonal antibodies against 5FITC have also been developed to detect the presence of toll-like receptor 4 (TLR4) on macrophages in vivo. 5FITC has also been used as a fluorescence probe for human immunoglobulin G (IgG) and erythrocytes in blood samples. Colloidal gold conjugates with 5FITC have been developed to measure the disulfide bonds in proteins such as human serum albumin.

5-Bromo-6-chloro-3-indoxyl phosphate, disodium salt monohydrate is a substrate used to visualize phosphatase activity via showing magenta colored end product when used with Nitroblue Tetrazolium.

4-Nitrophenyl beta-D-cellotrioside is a chromogenic substrate for cellulase. Upon hydrolysis, para-nitrophenol is released yielding a yellowish colour. 4-Nitrophenyl beta-D-cellotrioside is used in different applications such as the Biomass conversion research, paper industry applications

A fluorogenic MOSCERDAM® substrate targeting alpha-arabinofuranosidase

Chromogenic substrate for beta-glucanase

A substrate for beta-glucuronidase.

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.

Chromogenic substrate for ?-glucanase

Naphthol AS-BI N-acetyl-beta-D-glucosaminide is a fluorogenic substrate for the pararosaniline-glyoxalase (PG) reaction. This substrate is used to study enzymatic activity in two human isoenzymes of PG, PG I and PG II. The conversion of this substrate by PG I or PG II results in a color change from yellow to red. Naphthol AS-BI N-acetyl-beta-D-glucosaminide may be used as an experimental tool to measure the activity of these enzymes and to study the effect of inhibitors on their activity. It can also be used as a marker for studying enzyme transfer between cells.

Monohydrate; beta-glucuronidase substrate; yellow/pink color Beta-glucuronidase activity can be measured in a colorimetric assay with phenolphthalein beta-D-glucuronide as substrate. Starting assay conditions can include 30 µg of enzyme-containing material, 0.6 mM phenolphthalein beta-D-glucuronide in 100 mM sodium phosphate buffer in a total volume of 100 µL and monitoring absorbance changes at 540 nm. Temperature incubation can be used to promote the enzymatic activity (Galindo, 2012).

2-Nitrophenyl 2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-b-D-glucopyranoside is a chromogenic substrate used to detect and measure the activity of glycosidase enzymes. Glycosidases are enzymes that hydrolyze glycosidic bonds in carbohydrates, and are produced by many organisms. The substrate is cleaved by glycosidase to release a chromophore, which can be quantified via spectrophotometry analysis. 2-Nitrophenyl 2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-b-D-glucopyranoside is widely used in various applications, including enzyme kinetics studies, inhibitor screening, and the detection of NAGase activity in biological samples, such as serum, urine, and cell lysates. It is also used for the identification and characterization of glycosidases in microorganisms and the study of glycosylation-related processes in cells and tissues.

5-Bromo-6-chloro-3-indolyl sulfate potassium salt is a fluorogenic substrate that can be used for food testing. It is a conjugate of 5-bromo-6-chloroindolylsulfate and potassium, which is dissolved in buffer solution. This product can also be used as a chromogenic substrate in environmental testing, an enzyme substrate in diagnostics, and a ligand for bioluminescence. It has high purity and quality, and can be used for chemiluminescence or bioluminescence. The CAS number for this product is 6581-24-4.

Chromogenic substrate for ?-galactosidase

4-Nitrophenyl 2-acetamido-2-deoxy-a-D-glucopyranoside is a chromogenic pNP substrate specifically designed for the analysis of N-acetylglucosaminidase activity. Upon enzyme action, the substrate releases 4-nitrophenol, a yellow compound that can be detected by spectrophotometric methods, providing a reliable and sensitive means of quantifying enzyme activity. This versatile substrate is widely used in biochemical studies, disease diagnostics, and the enzyme production industry.

Ready-to-use chemiluminescence substrate solution for AP detection

5-Nitro-3-indolyl-b-D-galactopyranoside is a substrate for a number of enzymes, including beta-galactosidase, beta-glucuronidase, and beta-mannosidase. It is used as an enzyme substrate in diagnostics to measure the activity of these enzymes. 5N3IG was also used to study the effects of conjugation on enzyme activity. 5N3IG is available in high purity and high quality at CAS No.

5-Bromo-3-indolyl-a-D-galactopyranoside (5BIG) is a bioluminescent substrate that is used in various applications, including environmental testing, diagnostics, and the detection of bacterial activity. 5BIG is a ligand for luciferase enzymes and is used as a chromogenic substrate. It also has the ability to be conjugated with other molecules for use in enzyme substrates or fluorescence assays. 5BIG has been shown to be an effective fluorogenic substrate for enzymes such as alkaline phosphatase and beta-glucuronidase.

6,8-Difluoro-4-methylumbelliferyl 2-acetamido-3-O-(b-D-galactopyranosyl)-a-D-galactopyranoside is a chemiluminescent substrate that reacts with oxygen in the presence of luciferase to produce light. This product is an excellent replacement for luminol, especially in applications where aqueous solvents are not suitable. 6,8-Difluoro-4-methylumbelliferyl 2-acetamido-3-O-(b-D-galactopyranosyl)-a-D-galactopyranoside has been extensively tested and shown to be nonhazardous in a variety of different culture media. It is also nonhazardous to the environment, as it can be easily degraded by microorganisms.

2-O-(4-Nitrophenyl)-a-D-N-acetylneuraminic acid ammonium salt is a chromogenic substrate commonly used in the detection and quantification of sialidase activity. Sialidases are a family of enzymes that hydrolyze sialic acid residues from glycoconjugates. This substrate is used to detect bacterial and viral sialidases, which play an important role in pathogen infectivity. The ammonium salt form of the substrate is preferred for its solubility in water-based assay systems.

Aldol® 467 N-acetyl-beta-D-glucosaminide is a chromogenic and fluorogenic enzyme substrate used for detection of ?-N-acetyl-glucosaminidase activity. The colorless enzyme substrate is cleaved in the presence of ?-N-acetyl-glucosaminidase yielding development of a yellow product. In addition, green fluorescence is generated when a suitable matrix or Aldol® 355 fluorescence enhancer is present. Aldol® 467 N-acetyl-beta-D-glucosaminide amide can be used in enzymatic assays or in live bacterial cultures. Aldol® 467 N-acetyl-beta-D-glucosaminide is suitable for use in liquid and solid media, under aerobic as well as anaerobic conditions.

3-indolyl-N-methyl a-D-arabinopyranoside is a fluorogenic substrate for the detection of beta-galactosidase. It also has been used as a chromogenic substrate for the detection of beta-glucuronidase. This product can be conjugated to an enzyme and used in enzyme substrates for the detection of beta-galactosidase, beta-glucuronidase, and other enzymes. 3-indolyl-N-methyl a-D-arabinopyranoside is also commonly used in culture media to detect bacteria and yeast.