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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.

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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.

Keywords:
BCABradfordELISALowryProteinQuantitative immuno-PCR (qIPCR)

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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.