Horseradish peroxidase
The enzyme horseradish peroxidase (HRP), found in horseradish, is used extensively in molecular biology applications primarily for its ability to amplify a weak signal and increase detectability of a target molecule. For example, an antibody conjugated to HRP may be used to detect a small amount of a specific protein in a western blot. Here, the antibody provides the specificity to locate the protein of interest and the HRP enzyme, in the presence of a substrate, produces a detectable signal[1]. Horseradish peroxidase is also commonly used in techniques such as ELISA and Immunohistochemistry.
Horseradish peroxidase is ideal in many respects for these applications because it is smaller, more stable and less expensive than other popular alternatives such as alkaline phosphatase. It also has a high turnover rate that allows generation of strong signals in a relatively short time span.
Another example, "In recent years the technique of marking neurons with the enzyme horseradish peroxidase has become a major tool. In its brief history, this method has probably been used by more neurobiologists than have used the Golgi stain since its discovery in 1870."[2]
Substrates
Numerous substrates for the horseradish peroxidase enzyme have been described and commercialized to exploit the desirable features of HRP. These substrates fall into several distinct categories. HRP catalyzes the conversion of chromogenic substrates (e.g. TMB, DAB) into intensely colored molecules. In contrast, chemiluminescent substrates (e.g. SuperSignal, ECL) generate light when acted upon by HRP.
References
- ^ Chau YP, Lu KS (1995). "Investigation of the blood-ganglion barrier properties in rat sympathetic ganglia by using lanthanum ion and horseradish peroxidase as tracers". ACTA ANATOMICA (BASEL). 153 (2): 135–144. PMID 8560966.
- ^ Quoted in "Cell Marking with Horseradish Peroxidase" adapted from D. Purves and J.W. Lichtman, Principles of Neural Development. Sinauer Associates, Inc., Sunderland, 1985.
External links
- Horseradish+peroxidase at the U.S. National Library of Medicine Medical Subject Headings (MeSH)