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The ELISA Master Guide discusses how to perform enzyme-linked immunosorbent assays (ELISAs) and provides real-world insights for optimal liquid handling and tool selection.
The guide provides advice from PhD-level cellular and molecular biologists with extensive experience performing ELISAs on how researchers can:
- Choose the most appropriate ELISA set-up for their research question
- Carry out successful ELISAs with a range of sample types
- Improve precision and reproducibility
- Minimize the risk of inaccurate results
This guide is structured into four parts:
- An overview of the different types of ELISA (direct, indirect, sandwich, competitive and ELISpot assays
- Factors influencing reproducibility in ELISA protocols: subpar reagent and consumable quality, cross-contamination, pipetting techniques, cross-reactivity, matrix-effect, batch-to-batch reagents and data analysis. For each aspect, the paper provides practical tips for overcoming these challenges and discusses the different types of liquid handling equipment available.
- Detailed step-by-step instructions on how to perform an ELISA workflow, using a typical sandwich ELISA carried out in a 96-well plate as an example.
- A discussion of fundamental applications of ELISAs within different areas of research, diagnostics, biopharma, and biotech. This includes real-world examples of ELISA in medical diagnosis, food and environmental testing, drug discovery and development, and quality control.
