Posts Tagged ‘EnzymeLinked’
ENZYME-LINKED IMMUNOSORBANT ASSAY (ELISA): An entry from Gale’s World of Microbiology and Immunology
Last Updated on Thursday, 24 June 2010 02:55 Written by Administrator Thursday, 24 June 2010 02:55
ENZYME-LINKED IMMUNOSORBANT ASSAY (ELISA): An entry from Gale’s World of Microbiology and Immunology
This digital document is an article from World of Microbiology and Immunology, brought to you by Gale®, a part of Cengage Learning, a world leader in e-research and educational publishing for libraries, schools and businesses. The length of the article is 493 words. The article is delivered in HTML format and is available in your Amazon.com Digital Locker immediately after purchase. You can view it with any web browser. Covers the concepts, theories, discoveries, and pioneers in microbiology and immunology, using a mix of traditional academic and topical articles, this title addresses current ethical, legal, and social issues with special emphasis given to biological warfare and terrorism.
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Tags: assay, elisa, entry, EnzymeLinked, from, Gale's, Immunology, IMMUNOSORBANT, Microbiology, World | Posted under Elisa Assay Books | No Comments
Last Updated on Thursday, 24 June 2010 01:11 Written by Administrator Thursday, 24 June 2010 01:11
Question by heyheyhey: Which component is colored in ELISA (Enzyme-linked Immunosorbent Assay)?
also, In an ELISA used to detect antibody, which component is linked to the enzyme?
Best answer:
Answer by spasmodeus
The color comes from the chemical alteration of an enzyme substrate, such as 3,3-diaminobenzidine (DAB). Conversion of DAB by an enzyme like horseradish peroxidase (HRP) produces a brown color which can be detected using a spectrophotometer.
The way it typically works is a multiwell assay plate is coated with an antibody against the desired antigen for detection. Samples are loaded into the wells, and the antigen allowed to bind to the antibody coating the plate. Unbound material is washed away, and a secondary antibody conjugated to an enzyme (like HRP) is loaded into the well. This antibody binds to the antigen already immobilized in the well by the coating antibody, forming a so-called “sandwich”. Finally, after unbound secondary antibody is washed away, the enzyme substrate (such as DAB) is introduced. The amount of enzyme still present in the well is directly proportional to the amount of antigen captured in the well and cross-linked to the enzyme by the secondary antibody. The amount of colored enzyme product is also proportional to the amount of enzyme. So, one can compare the amount of product produced from well to well over time, and from this value quantitate the amount of antigen present in a particular sample.
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Tags: assay, colored, component, elisa, EnzymeLinked, Immunosorbent | Posted under Elisa Assay Questions | No Comments
Last Updated on Tuesday, 22 June 2010 04:57 Written by Administrator Tuesday, 22 June 2010 04:57
Strategies for direct attachment of hapten to a polystyrene support for applications in enzyme-linked immunosorbent assay (ELISA) [An article from: Analytica Chimica Acta]
This digital document is a journal article from Analytica Chimica Acta, published by Elsevier in 2004. The article is delivered in HTML format and is available in your Amazon.com Media Library immediately after purchase. You can view it with any web browser.
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A new method describing direct attachment of carboxylated haptens on a polystyrene support, using 3-aminopropyltriethoxysilane (3-APTES) as a linker, is reported. The hapten coated polystyrene support showed excellent stability as a function of the buffer pH and reaction time, and was successfully used to demonstrate its application in enzyme-linked immunosorbent assay (ELISA).
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Tags: Acta, Analytica, applications, article, assay, attachment, Chimica, direct, elisa, EnzymeLinked, from, hapten, Immunosorbent, polystyrene, Strategies, support | Posted under Elisa Assay Books | No Comments
Last Updated on Tuesday, 22 June 2010 11:28 Written by Administrator Tuesday, 22 June 2010 11:28
A rapid and sensitive chemiluminescence enzyme-linked immunosorbent assay for the determination of fumonisin B”1 in food samples [An article from: Analytica Chimica Acta]
This digital document is a journal article from Analytica Chimica Acta, published by Elsevier in 2006. The article is delivered in HTML format and is available in your Amazon.com Media Library immediately after purchase. You can view it with any web browser.
Description:
An enzyme-linked immunosorbent assay (ELISA) based on polyclonal antibody with enhanced chemiluminescent (ECL) detection of fumonisin B”1 (FB”1) in food samples has been developed. Assay conditions, including concentrations of antibody and enzyme conjugate, competition time and so on, were optimized. The effects of pH and two different organic solvents were investigated. The optimized ECL-ELISA system allowed FB”1 determination in a linear working range of 0.14-0.9@mgL^-^1 with IC”5″0 value of 0.32@mgL^-^1 and a limit of detection of 0.09@mgL^-^1. The ECL-ELISA was about 10 times more sensitive and about 30% time less than that of colorimetric ELISA using the same antibody and HRP-conjugate. Good recoveries with spiked food samples were obtained, and the results correlated well with those obtained using conventional direct competition ELISA assay and HPLC method, which indicated that ECL-ELISA was capable of being applied for the specific detection and routine monitoring of FB”1 in food samples.
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Tags: Acta, Analytica, article, assay, chemiluminescence, Chimica, determination, EnzymeLinked, food, from, fumonisin, Immunosorbent, rapid, samples, sensitive | Posted under Elisa Assay Books | No Comments