Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023:2699:305-347.
doi: 10.1007/978-1-0716-3362-5_16.

Protein Quantitation and Analysis of Purity

Eva M Campion  1 , Dermot Walls  2 , Sinéad T Loughran  3
Affiliations

Protein Quantitation and Analysis of Purity

Eva M Campion et al. Methods Mol Biol. 2023.

Abstract

The accurate quantitation of proteins and an analysis of their purity is essential in numerous areas of scientific research and is a critical factor in many clinical applications. The large number and variety of techniques employed for this purpose is therefore not surprising. The selection of a suitable assay is dependent on such factors as the level of sensitivity required, the presence of interfering agents, and the composition of the protein itself. In this chapter, protocols for the most commonly used protein determination methodologies are outlined, including an overview of the highly sensitive real-time quantitative immuno-polymerase chain reaction assay. In addition, an approach to validate the UV protein absorption assay is outlined, which can be applied to any procedure for method validation.

Keywords: BCA; Bradford; ELISA; Lowry; Protein; Quantitative immuno-PCR (qIPCR).

PubMed Disclaimer

References

    1. Rosenburg IM (2005) Getting started with protein purification. In: Protein analysis and purification: Benchtop techniques, 2nd edn. Birkhäuser, Boston, pp 118–152
    1. Goldring JPD (2019) Measuring protein concentration with absorbance, Lowry, Bradford Coomassie Blue, or the Smith Bicinchoninic acid assay before electrophoresis. Methods Mol Biol 1855:31–39. https://doi.org/10.1007/978-1-4939-8793-1_3 - DOI - PubMed
    1. Kielkopf CL, Bauer W, Urbatsch IL (2020) Methods for measuring the concentrations of proteins. Cold Spring Harb Protoc 2020(4):102277. https://doi.org/10.1101/pdb.top102277.15 - DOI
    1. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein--dye binding. Anal Biochem 72:248–254 - DOI - PubMed
    1. Kielkopf CL, Bauer W, Urbatsch IL (2020) Bradford assay for determining protein concentration. Cold Spring Harb Protoc 2020(4):102269. https://doi.org/10.1101/pdb.prot102269 - DOI

MeSH terms

LinkOut - more resources