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The enzyme horseradish peroxidase (HRP), found in the roots of horseradish, is used extensively in biochemistry applications. It is a metalloenzyme with many isoforms, of which the most studied type is C. It catalyzes the oxidation of various organic substrates by hydrogen peroxide.
Peroxidase can be used for treatment of industrial waste waters. For example, phenols, which are important pollutants, can be removed by enzyme-catalyzed polymerization using horseradish peroxidase. Thus phenols are oxidized to phenoxy radicals, which participate in reactions where polymers and oligomers are produced that are less toxic than ...
3,3′,5,5′-Tetramethylbenzidine or TMB is a chromogenic substrate used in staining procedures in immunohistochemistry as well as being a visualising reagent used in enzyme-linked immunosorbent assays . [1] TMB is a white solid that forms a pale blue-green liquid in solution with ethyl acetate.
A common use for it is in the enzyme-linked immunosorbent assay (ELISA) to detect the binding of molecules to each other. It is commonly used as a substrate with hydrogen peroxide for a peroxidase enzyme (such as horseradish peroxidase) or alone with blue multicopper oxidase enzymes (such as laccase or bilirubin oxidase).
A suitable alternative to radioimmunoassay would substitute a nonradioactive signal in place of the radioactive signal. When enzymes (such as horseradish peroxidase) react with appropriate substrates (such as ABTS or TMB), a change in color occurs, which is used as a signal. However, the signal has to be associated with the presence of antibody ...
Some examples of substrates used (also known as chromogens) are AEC (3-Amino-9-EthylCarbazole), or DAB (3,3'-Diaminobenzidine). Use of one of these reagents after exposure to the necessary enzyme (e.g., horseradish peroxidase conjugated to an antibody reagent) produces a positive immunoreaction product.
This enzyme complex then catalyzes the conversion of the enhanced chemiluminescent substrate into a sensitized reagent in the vicinity of the molecule of interest, which on further oxidation by hydrogen peroxide, produces a triplet (excited) carbonyl, which emits light when it decays to the singlet carbonyl. Enhanced chemiluminescence allows ...
The surrounding solution would end up containing the enzyme lipase B with the glucose molecules containing four acetyl groups as the substrate. [6] The resulting glucose would cross the membrane, become oxidized, and the horseradish peroxidase would convert the sample substrate ABTS (2,2’-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)) into ...