Enzyme Substrates

AquaSpark® Peroxide Probe is a dioxetane-based substrate for the luminescence detection of hydrogen peroxide and other peroxides in biological samples. AquaSpark® Peroxide Probe is the most sensitive probe for H2O2 detection available. It is also an extremely versatile probe for oxidase detection applicable in a wide range of biocompatible buffers. For example it can be used for glucose oxidase detection andfor the quantitation of glucose levels (when coupled to the glucoseoxidase activity in an assay). In addition, AquaSpark® Peroxide Probe is used for the quantification of other biologically relevant compounds(e.g. amino acids) using the corresponding peroxide releasing oxidases.

5-Bromo-4-chloro-3-indoxyl phosphate, p-toluidine salt - micronized is a chemiluminescent substrate used in diagnostic assays. It is an enzyme substrate that is commonly used to detect peroxidase activity and has been shown to be effective in detecting various types of bacteria. This product is a high quality chemical with a purity of at least 98% and has been tested for environmental safety. 5-Bromo-4-chloro-3-indoxyl phosphate, p-toluidine salt - micronized can be used as a fluorogenic substrate for the detection of proteases and aminopeptidases.

5-Bromo-4-Chloro-3-Indolyl b-D-Mannopyranoside, also known as X-Man, is an enzyme substrate commonly used for detecting mannosidase enzymes. Upon hydrolysis by the enzyme, it produces a blue-green colored compound that can be detected visually or measured spectrophotometrically. This substrate is useful in characterizing the activity of mannosidases involved in glycoprotein processing and quality control.

Chromogenic substrate for beta-glucuronidase (GUS) yielding blue precipitate upon cleavage. Used for identification of beta-glucuronidase. X-beta-D-glucuronide has been reported as chromogenic substrate in a number of chromogenic E. coli/ coliform culture media. About 95% of common E. coli possess beta-D-glucuronidase activity. Such chromogenic culture media are used for detection and enumeration of E. coli in food samples such as meat, dairy products and shellfish and have clinical applications in the assessment of urinary infections. It has gained international acceptance as highly accurate test for the presence of E. coli in drinking water samples. It is also used for the detection of the expression of GUS (E. coli beta-glucuronidase) gene fusion marker constructs in plants. The sodium salt has advantages over the cyclohexylammonium salt as being non-electrostatic and possessing a naturally-occurring cation.

A fluorogenic substrate for the detection of lysosomal enzyme N-acetyl-α-D-galactopyranosiaminidase (NAGA), which is driving the pathogenesis of Schindler disease. Also substrate for endo-α-N-acetylgalactosaminidase (O-glycosidase), which cleaves O-linked core 1 and core 3 disaccharides from glycoproteins.

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Chromogenic substrate targeting alpha-L-arabinofuranosidase

5-Bromo-4-chloro-3-indoxyl-1-acetate is a chromogenic substrate used to detect the specific enzymatic activity of esterase. After cleavage, 5-bromo-4-chloro-indoxyl is released, resulting in a blue to blue-green color change in bacterial colonies or media. 5-Bromo-4-chloro-3-indoxyl-1-acetate is used in bacterial esterase activity assays (e.g. Pseudomonas spp.).

Aldol® 495 nonanoate is a chromogenic and fluorogenic enzyme substrate used for detection of lipase activity. The colorless enzyme substrate is cleaved by C9 esterases, yielding development of an orange coloration. In addition, green fluorescence is generated when Aldol® 355 fluorescence enhancer or a suitable matrix is present. Aldol® 495 nonanoate can be used in enzyme assays and live bacterial cultures. Aldol® 495 nonanoate is suitable for use in liquid and solid media, under aerobic as well as anaerobic conditions.

6-Chloro-3-indoxyl-beta-D-galactopyranoside is a chromogenic substrate for beta-galactosidase that yields a salmon or rose-coloured precipitate and hence is also known as Salmon-Gal. It can be used in conjunction with isopropyl-beta-D-thiogalactopyranoside (IPTG) as an alternative to the chromogenic substrate 5-bromo-4-chloro-3-indolyl beta-D-galactopyranoside (X-Gal) for detection of beta-galactosidase activity in bacterial colonies in colorimetric assays. Where beta-galactosidase expressed from a reporter gene in molecular biology experiments, recombinant colonies on growth plates are salmon-coloured while non-recombinants appear as white. Use of nitroblue tetrazolium salt (NBT) with Salmon-Gal may provide faster results, yielding a clear, purple precipitate, than for Salmon-Gal alone. In conjunction with 5-bromo-4-chloro-3-indolyl-beta-D-glucoside (X-Glc), Salmon-Gal is useful for simultaneous detection of GUS and Lac activities in the same medium.

AquaSpark® H2S/-SH is a chemiluminescent probe for the detection of sulfides and thiols. The chemiluminescence obtained through sulfide is 3-5 fold higher than chemiluminescence obtained through thiols. AquaSpark® H2S/-SH probe was used in a study identifying antibiotic-resistant bacterial strains by detecting the beta-lactamase-driven hydrolysis of beta-lactam antibiotics, where hydrogen sulfide is a biodegradation metabolite (Popat Gholap et al, 2021).

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

4-Nitrophenyl O-(N-acetyl-a-neuraminosyl)-(2-3)-b-D-galactopyranosyl-(1-4)-2-acetamido-2-deoxybDglucopyranoside is a fluorescent compound that has been used as a substrate in enzyme assays. It has been shown to be an effective ligand for the enzyme beta galactosidase, which hydrolyzes the substrate to produce 4nitrophenol and o-(Nacetyl)a-neuraminic acid. This product is CAS No. 501427-92-5 and can be used as a high purity, high quality reagent for conjugation.

4-Methylumbelliferyl-alpha-L-arabinofuranoside is a nucleic acid molecule that has an average diameter of 10 nm. It is used as a tracer in electron microscopy to detect the presence of nucleic acid molecules in cells. This compound is also used to study the interactions between DNA and RNA molecules by labeling the DNA strand with 4-methylumbelliferyl-alpha-L-arabinofuranoside, which contains an alpha-(2,4,6) arabinofuranosyl group. The recombinant expression of this compound can be carried out in corynebacterium glutamicum, and it has a bulk density that ranges from 1.7 to 2.1 g/cm3 and an average particle diameter ranging from 0.5 to 1 micron. It is not soluble in water or alcohol but is soluble in acetone or chloroform.

Applications A novel fluorigenic substrate for the determination of Biotinidase activity. References Kumugi, S., et al.: Chemistry Letters, 391 (1997)

Fluorogenic substrate for esterase yielding a blue fluorescent solution upon cleavage.

This Clips-O substrate rac-7-(3',4'-Dihydroxybutyloxy)coumarin-4'-phosphate allows the detection of single and dual enzymatic activity (phosphatase and oxidation of 4-amino-3-hydroxybutyloxy spacer either enzymatically or by exposure to sodium periodate) with a single substrate resulting in a fluorescent signal. The main virtues of Clips-O substrates are an increased stability towards non-enzymatic hydrolysis that results in a reduced background and high sensitivity.

Resorufin phosphate disodium is a fluorogenic substrate for phosphatase exhibiting a red color after cleavage that can be measured at ~ 585nm.

5-Bromo-4-chloro-3-indolyl b-D-lactopyranoside is a chromogenic substrate used to detect the specific enzymatic activity of beta-galactosidase. After cleavage, 5-bromo-4-chloro-indoxyl is released, resulting in a blue to blue-green color change in bacterial colonies or media. 5-Bromo-4-chloro-3-indolyl b-D-lactopyranoside is used in lactose intolerance screening.

2-O-(4-Nitrophenyl)-a-D-N-acetylneuraminic acid, also known as p-nitrophenyl N-acetylneuraminic acid, is a chromogenic substrate specifically designed for the detection and quantification of neuraminidase (sialidase) activity. Upon cleavage by neuraminidase, it releases a yellow-colored product, 4-nitrophenol, which can be easily monitored spectrophotometrically at 405 nm. This substrate is widely used in various applications, including enzyme kinetics studies, inhibitor screening, and the determination of neuraminidase activity in biological samples such as viruses, bacteria, and cell lysates. It is particularly useful for studying the enzymatic properties of neuraminidases from different sources and for the development of novel antiviral and antibacterial agents targeting neuraminidase activity.

2'-(4-Methylumbelliferyl)-a-D-N-glycolylneuraminic acid is a fluorogenic substrate that is used for enzymatic labeling of glycolipids. It can be used as a diagnostic or food testing agent, and also has applications in environmental testing. The CAS number for this product is 1434265-32-3.

4-Nitrophenyl 4,6-ethylidene-?-D-maltoheptaoside is a substrate used in a reference measurement method for ?-amylase activity in the diagnosis of acute pancreatitis, approved by the International Federation of Clinical Chemistry (IFCC). The substrate is hydrolysed by serum ?-amylase into shorter 4-nitrophenylated oligosaccharides and their free counterparts. These reaction products are channeled into an indicator reaction with ?-glucosidase and are degraded into glucose and the chromophore, 4-nitrophenol.

4-Methylumbelliferyl 6-O-(N-acetylneuraminyl)-b-D-glucopyranoside is a fluorescent substrate for the enzyme β-D-galactosidase. It is used in the detection of β-D-galactosidase activity in the diagnosis of various diseases, including celiac disease and AIDS. 4-Methylumbelliferyl 6-O-(N-acetylneuraminyl)-b-D-glucopyranoside can also be used as a chromogenic or fluorogenic substrate for chemiluminescent detection of β-D-galactosidase activity. The conjugate with horseradish peroxidase can be used to detect hydrogen peroxide in water samples, such as drinking water, wastewater, and surface waters. 4MUG has been shown to have high purity and is CAS No. 78761-35-4.

Chromogenic substrate targeting alpha-D-galactosidase

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.

4-Nitrophenyl 3,4-O-isopropylidene-a-D-galactopyranoside is a chemiluminescent substrate for detecting beta-galactosidase activity in bacteria. This product is a high purity, high quality compound that can be used as a culture media and environmental testing agent. It has been shown to be effective in diagnostics, chromogenic substrate, and enzyme substrate reactions. 4NPPG has been shown to be a fluorogenic substrate for beta-galactosidase assay and can be used as food testing agent with conjugate.

4-Nitrophenyl 2-acetamido-3-O-acetyl-4,6-O-benzylidene-2-deoxy-aDglucopyranoside is a fluorogenic substrate that can be used for the detection of β-glucosidase activity. It is a water soluble substrate with an extinction coefficient of approximately 10,000 M−1 cm−1 at 420 nm. 4NPAABGDG is a chromogenic substrate that can be used to detect β-galactosidase activity. It has an extinction coefficient of approximately 1,700 M−1 cm−1 at 420 nm. The CAS number for 4NPAABGDG is 23262-56-8 and it is sold by Sigma Aldrich under catalog number L4959.

4-Methylumbelliferyl 3,4,6-tri-O-acetyl-b-D-galactopyranoside is a fluorescent compound that can be used as a substrate for the following enzymes: β-galactosidase, α-galactosidase, and β-glucuronidase. This product is used in diagnostics to measure the activity of these enzymes. It can also be used to detect bacteria in culture media and food testing. 4MUF3,4,6TABG is a high quality product that has been shown to have a high purity level by liquid chromatography. CAS number: 61440-03-2

Fluorogenic substrate for ?-galactosidase

A substrate for visualising alkaline phosphatase activity. It produces a purple insoluble end product that is detected visually, when used together with nitroblue tetrazolium. The substrate system is versatile and has functions in a variety of applications, including Northern, Southern, and Western blotting, in situ hybridization, ELISAs, microarray and immunohistochemistry. DMF soluble.

Fluorogenic substrate for aminopeptidase

Histamine (Scombrotoxin) fish poisoning is the most frequent cause of fish poisoning incidents today. Cymit Quimica's BCM®; Histamine-producingbacteria Broth (C-0450) is a growth medium for the detection of histamine-producing bacteria in food and water samples. The medium contains an indicator that turns orange to red due to metabolites released by histidine decarboxylase.

4-Nitrophenyl 6-O-(alpha-D-glucopyranosyl)-beta-D-glucopyranoside is a chromogenic substrate for beta-glucosidase. Upon hydrolysis, para-nitrophenol is released yielding a yellowish colour. 4-Nitrophenyl 6-O-(alpha-D-glucopyranosyl)-beta-D-glucopyranoside is used in different applications such as the Cellulose metabolism research, biofuel production

2-Chloro-4-nitrophenyl b-D-cellobioside (CNPG) is a chromogenic enzyme substrate that is commonly used to detect and measure the activity of β-glucosidases. CNPG is cleaved by the enzyme into the chromogenic product, which can be detected spectrophotometrically. β-glucosidases are important enzymes in the breakdown of cellulose, and CNPG is a widely used substrate for the detection and quantification of these enzymes in various research studies.

AquaSpark™ α-D-galactoside is a dioxetane-based substrate for the luminescence detection of α-galactosidase activity. It is the first single-compound chemiluminescent substrate commercially available for the detection of α-galactosidase activity. α-galactosidase assays are used in molecular biology to monitor the transcriptional activity of the GLA gene that is correlated with metabolic diseases or gene mutations. In microbiology the α-galactosidase activity is used to detect specific bacteria such as α-galactosidase positive Salmonella strains (Salmonella Enteritidis RKI 05/07992).

6-Chloro-1H-indol-3-yl octanoate is a fluorogenic substrate that can be used to detect enzymes such as phosphatases, lipases, esterases, and proteases. 6-Chloro-1H-indol-3-yl octanoate is a ligand for metal ions such as Cu(II), Ni(II) and Zn(II). This product has high purity and quality. It is an enzyme substrate for many enzymes such as phosphatases, lipases, esterases, and proteases. This product can be used in diagnostics of various substances including food testing, environmental testing and staining. 6CIO is a chromogenic substrate that can be used to detect bioluminescence reactions. 6CIO is CAS No. 159954-35-5.

2-Nitrophenyl 2,3-di-O-acetyl-b-D-xylopyranoside is a chromogenic substrate used to assay the activity of xylosidase enzymes. It is hydrolyzed by xylosidase to yield a yellow-colored product, 2-nitrophenol, which absorbs light at 405 and 410 nm. This substrate is used in research to study the function of xylosidase enzymes and their role in various biological processes. It is also used in diagnostic tests for the detection of xylosidase activity in different microorganisms, including bacteria and fungi.

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4-Methylumbelliferyl b-D-mannopyranoside is a fluorescent probe that can be used to measure the activity of the enzyme β-glycosidase. It is synthesized from 4-methylumbelliferone, which reacts with b-D-mannopyranosyl chloride and pyridine, followed by glycosylation with glucal. The maximum absorption of this compound occurs at a wavelength of 400 nm in an acidic solution. The pH optimum for this compound is 5.5, so it should not be used in tissues or organs where the pH is greater than 7.4. This product has been found to have no adverse effects on animals and does not react with other enzymes, making it suitable for use as a biochemical marker for liver cells or tissues.

D-Alanine 7-amido-4-methylcoumarin trifluoroacetate salt is a chromogenic substrate for the enzyme Beta-galactosidase. It is used as a diagnostic tool in the detection of certain bacteria, such as Enterobacteriaceae and Clostridium perfringens. It can also be used to detect the presence of beta galactosidase in food products and environmental samples. D-Alanine 7-amido-4-methylcoumarin trifluoroacetate salt has an applications in chemiluminescence, bioluminescence, and fluorescence. This product is not intended for use as a drug or food additive.

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This product is a fluorogenic substrate that reacts with the enzyme beta-galactosidase to produce a fluorescent product. It can be used in food testing, environmental testing, and diagnostics. The CAS number for this product is 609-33-2. This product is available in high purity and high quality.