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Questions and Answers
What is added after the well is washed in a sandwich ELISA?
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 does the second enzyme-linked antibody react with in a sandwich ELISA?
What is the purpose of adding substrate 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?
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?
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?
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?
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?
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?
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?
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?
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?
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.
