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Questions and Answers
What is added after the well is washed in a sandwich ELISA?
What does the second enzyme-linked antibody react with in a sandwich ELISA?
What is the purpose of adding substrate in a sandwich ELISA?
In a sandwich ELISA, what is the purpose of immobilizing the antibody on the microtiter well?
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Why is a second enzyme-linked antibody specific for a different epitope on the antigen used in a sandwich ELISA?
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In a sandwich ELISA, what is immobilized on the microtiter well?
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What is the significance of adding substrate in a sandwich ELISA?
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After washing the microtiter well in a sandwich ELISA, what is the purpose of adding a second enzyme-linked antibody?
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In competitive ELISA, what is the purpose of incubating antibody in solution with a sample containing antigen?
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What is the function of the enzyme-conjugated secondary antibody (Ab2) in competitive ELISA?
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Why does the absorbance decrease with an increase in the concentration of antigen in competitive ELISA?
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What role does the antigen-coated microtiter well play in competitive ELISA?
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Study Notes
Sandwich ELISA
- After washing, a second enzyme-linked antibody is added to bind to the antigen.
- The second enzyme-linked antibody reacts with a different epitope on the antigen, ensuring specificity.
- Adding substrate is crucial as it reacts with the enzyme to produce a measurable signal, usually a color change.
- Immobilizing the antibody on the microtiter well captures the target antigen, facilitating its detection.
- A second enzyme-linked antibody specific for a different epitope enhances the assay's sensitivity and specificity.
- The microtiter well is coated with the capture antibody, which binds the target antigen.
- The added substrate's significance lies in generating a detectable signal proportional to the antigen amount.
Competitive ELISA
- Incubating antibody in solution with sample antigen allows the antigen in the sample to compete with the immobilized antigen.
- The enzyme-conjugated secondary antibody (Ab2) in competitive ELISA binds to any unoccupied antibody, facilitating quantification.
- Absorbance decreases with increased antigen concentration because more antigen competes for binding, reducing the signal.
- The antigen-coated microtiter well serves as a platform that binds the competing antibodies, essential for the assay's detection process.
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Description
Learn about the Sandwich ELISA technique where the antibody is immobilized on a microtiter well, and a sample containing antigen is added to react with the immobilized antibody. This quiz covers the process and steps involved in this particular ELISA technique.
