Team:WLC-Milwaukee/Background
BACKGROUND
Introduction
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Water Contamination and Testing (United States and Global)
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Lambda Phage
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Horse Radish Peroxidase
Horseradish peroxidase (HRP) is an enzyme used in molecular biology. It can be used in various biological methods, such as ELISA, Western Blotting, Southern Blotting, and EMSA [1]. HRP’s colorimetric and chemiluminescent properties make it a useful agent in many tests. HRP acts as a catalyst in redox reactions [3]. A luminol and H2O2 solution can release light for a very long time, but when HRP is added, it usually reaches it maximum light emission between 1 and 5 minutes [4]. This makes it a useful detection method. HRP can be used in more qualitative methods as well, since it induces changes in color when used with chromogenic assays. Commonly used substrates for this type of detection are 3,3',5,5'-tetramethylbenzidine (TMB), 2,2' -azino-di-[3-ethylbenzthiazoline-6-sulfonic acid] (ABTS), and diaminobenzidine (DAB) [3,4]. HRP when added to TMB, for example, yields a 2-electron oxidation product which yields a blue color. HRP is also up and coming in the medical field. When combined with other agents, it is heavily reactive toward tumor cells in humans [2]. It also serves as a functional tracer. HRP can be injected into the bloodstream and then identified at different places in the pathway using the DAB/H2O2 reaction [4]. This makes HRP very useful for immunocytochemistry and electron microscopy [4].
[1] https://www.sigmaaldrich.com/life-science/biochemicals/biochemical-products.html?TablePage=15552120 [2]https://chem.libretexts.org/Textbook_Maps/Biological_Chemistry/Catalysts/Case_Studies/Horseradish_Peroxidase [3] http://www.bio-rad.com/featured/en/hrp-substrate.html [4] https://www.sciencedirect.com/topics/neuroscience/horseradish-peroxidase [5] http://www.jbc.org/content/257/7/3669.full.pdf
