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Questions and Answers

Which reagent should come first in a non-competitive immunoassay?

Anti-cortisol HRP conjugated Ab

Analyte (cortisol) (correct)

HRP

TMB substrate

What role does a blank serve in an immunoassay?

It contains the analyte of interest.

It represents the maximum signal obtainable.

It provides a control for specific binding.

It helps to correct the absorbances of standards and unknowns. (correct)

In a competitive immunoassay, what is represented by %B/B0?

Percentage of absorbance of each sample relative to the blank. (correct)

Percentage of blank binding.

Total absorbance of the sample.

Percentage of analyte bound to antibody.

What is the correct formula to calculate % Ab binding in a competitive assay?

<p>% Ab binding = (100 - %B/B0)</p> Signup and view all the answers

Which statement correctly describes a competitive immunoassay procedure?

<p>The analyte competes directly with the labeled antigen for binding to the antibody.</p> Signup and view all the answers

Study Notes

Data Handling – Part 3

Non-competitive Immunoassay Reagent Order: TMB substrate, HRP, Anti-cortisol HRP conjugated Ab, Analyte (cortisol), Anti-cortisol Ab

Competitive Immunoassay Reagent Order

Competitive Immunoassay Reagent Order: Labelled Ag (cortisol), Analyte Ag (cortisol), HRP conjugated Ab, Anti-cortisol Ab, TMB substrate

Blank Definition

Blank Definition: A sample lacking analyte, used to determine non-specific binding in an assay.
It corrects for background absorbances of standards and unknowns.

Non-competitive Assay Calculations

Non-competitive Assay Calculations: Subtract the blank absorbance from all sample absorbances (Sample Abs - Blank Abs)

Competitive Assay Calculations

Competitive Assay Calculation Formula: %B/B0 = (Absorbance of sample / Absorbance of blank) x 100
Calculate percentage Ab binding using the following formula: % Ab binding = (100 - %B/B0)

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Description

This quiz covers essential concepts in non-competitive and competitive immunoassays, including reagent orders and calculation methods. It highlights the importance of blank definitions and how to correct for background absorbances. Test your knowledge on these critical assays and their applications.