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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) (C)</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. (A)</p> Signup and view all the answers

Flashcards

Blank

A sample without the analyte being measured, used to determine non-specific binding in the assay.

Competitive Immunoassay

A type of immunoassay where the analyte and a labeled analyte compete for binding to a limited amount of antibody.

Non-Competitive Immunoassay

A type of immunoassay where the analyte binds directly to an antibody, and the signal is directly proportional to the analyte concentration.

Standard Curve

A standard curve used to determine the analyte concentration in unknown samples. It plots known analyte concentrations against their corresponding absorbance values.

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TMB Substrate

A reagent that reacts with an enzyme to produce a measurable signal. It's used to visualize the antibody-antigen complex.

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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.