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Sandwich ELISA

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Learn about the reagents and steps involved in an ELISA, focusing on sandwich ELISAs. Review the protocol order and understand the relevant controls used in an ELISA.

Learning Outcomes

  • Learn about the reagents and steps involved in an ELISA
  • Answer questions about the theory of the ELISA
  • Familiarise yourself with the relevant controls used in an ELISA
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Concepts Principles and Theories

  • How antibodies can be used to detect the presence of an analyte within a sample
  • What is happening on a molecular level during each step of the ELISA 
  • What it means to immobilise the antigen and why this is necessary 
  • What is meant by a colorimetric reaction 
  • How the results of an ELISA can be used to estimate the concentration of an analyte in a sample

Using Instruments and Equipment

  • What are the core steps and components of an ELISA 
  • How to immobilise the antigen 
  • How to probe for the presence of antigen 
  • How to detect bound antigen

Obtaining and Recording Data

  • Why including blocking and wash steps increases the sensitivity of the assay 
  • Why samples, standards and controls should be assayed in duplicate or triplicate

Designing and Planning Experiments

  • What controls should be assayed alongside samples in an ELISA

Learn more about our approach to skills
PGR student, School of Pharmacy

"I accessed the Sandwich-ELISA resource from the PGR-Biosciences-Lab-Shots library as face-to-face training was to be avoided during Covid-19-restrictions. As a result, I felt much more confident when discussing the technique with my supervisor and undertaking ELISAs for the first time on my own in the lab."



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