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Protein Quantitation and Analysis of Purity
Eva M Campion1, Dermot Walls2, Sinéad T Loughran3
1Department of Life Science, Faculty of Science, Atlantic Technological University, Ash Lane, Sligo, Ireland. eva.campion@atu.ie.
Methods in Molecular Biology (Clifton, N.J.)
|August 30, 2023
Summary
Accurate protein quantitation and purity analysis are vital in research and clinical settings. This chapter details common protein determination methods, including real-time quantitative immuno-polymerase chain reaction (PCR) and UV absorption assay validation.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Molecular Biology
Background:
- Accurate protein quantitation and purity analysis are crucial for scientific research and clinical applications.
- A wide array of techniques exists for protein determination, necessitating careful selection based on specific experimental needs.
Purpose of the Study:
- To outline protocols for commonly used protein determination methodologies.
- To provide an overview of the highly sensitive real-time quantitative immuno-polymerase chain reaction (PCR) assay.
- To present an approach for validating the UV protein absorption assay.
Main Methods:
- Review and summarization of established protein determination techniques.
- Detailed protocol for real-time quantitative immuno-PCR.
- Methodology for validating UV protein absorption assays.
Main Results:
- Protocols for standard protein determination methods are provided.
- The application of real-time quantitative immuno-PCR for sensitive protein analysis is described.
- A framework for validating UV protein absorption assays is presented.
Conclusions:
- The chapter offers a comprehensive guide to protein quantitation and purity analysis methods.
- Selection of appropriate protein assays depends on sensitivity, interfering agents, and protein composition.
- Method validation, including for UV absorption assays, is essential for reliable results.
