BIOT6002 Lecture 15 Data Handling 3 PDF
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Uploaded by ClearerSaxhorn1261
Munster Technological University
Dr. Caroline A. Browne
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Summary
This document appears to be lecture notes covering immunoassay techniques and data handling. It outlines the procedures for non-competitive and competitive immunoassays. It also defines a blank and describes how it is used for calculating absorbance values.
Full Transcript
Data Handling – part 3 BIOT6002: Lecture 15 Lecturer: Dr. Caroline A. Browne Place these reagents in the correct order for a non-competitive immunoassay TMB substrate HRP Anti-cortisol HRP conjugated Ab Analyte (cortisol) Anti-cortisol Ab Place these reagents in the correct order for a...
Data Handling – part 3 BIOT6002: Lecture 15 Lecturer: Dr. Caroline A. Browne Place these reagents in the correct order for a non-competitive immunoassay TMB substrate HRP Anti-cortisol HRP conjugated Ab Analyte (cortisol) Anti-cortisol Ab Place these reagents in the correct order for a competitive immunoassay Labelled Ag (cortisol) Analyte Ag (cortisol) HRP conjugated Ab Anti-cortisol Ab TMB substrate What is a blank? A sample that does not contain any analyte can be referred as a blank. It shows the non-specific binding of the assay It is used to correct the absorbances of standards and unknowns. Non-competitive Assays Blank is subtracted from all absorbance values Sample Abs – Blank Abs Competitive immunoassays Calculate %B/B0 (% of Ab bound to HSA) for each sample by expressing the absorbance of each standard as a percentage of the absorbance of the blank (B0) %B/B0 = (Absorbance of sample / Absorbance of blank) x 100 Competitive assay Calculate the percentage Ab binding using the following formula: % Ab binding = (100 – %B/B0)
