Abstract
Although a lot of new methods for protein concentration determination have been developed and established the last years, the amino acid analysis has still this relevance within proteomics for multiple reasons especially in the quantitative protein analysis. Amino acid analysis enables indirectly both the protein and peptide concentration determination which are essential for using the same amounts for comparative quantitative experiments. Moreover, the quantity and quality of synthetic peptides can be verified. The method itself is robust in comparison with colorimetric assays, especially when detergents or chaotropes are present in the sample buffer. Furthermore, it is highly sensitive. Nevertheless, amino acid analysis needs a certain experience to be set up and is time-consuming compared to other protein concentration determination techniques.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Lowry OH, Rosebrough NJ, Farr AL et al (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193(1):265–275
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
Gornall AG, Bardawall CJ, David MM (1949) Determination of serum proteins by means of the biuret reaction. J Biol Chem 177(2):751–766
Guntermann A, Steinbach S, Serschnitzki B et al (2019) Human tear fluid proteome dataset for usage as a spectral library and for protein modeling. Data Brief 23:103,742
Acknowledgments
This work was supported by the Deutsche Parkinson Gesellschaft, the Bundesministerium für Bildung und Forschung, Germany (WTZ mit Brasilien, FKZ 01DN14023), the HUPO Brain Proteome Project, and P.U.R.E. (Protein Research Unit Ruhr within Europe) and ProDi (Center for protein diagnostics), Ministry of Innovation, Science and Research of North-Rhine Westphalia, Germany, a federal German state, FoRUM (F781-13).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
May, C., Serschnitzki, B., Marcus, K. (2021). Good Old-Fashioned Protein Concentration Determination by Amino Acid Analysis. In: Marcus, K., Eisenacher, M., Sitek, B. (eds) Quantitative Methods in Proteomics. Methods in Molecular Biology, vol 2228. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1024-4_2
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1024-4_2
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1023-7
Online ISBN: 978-1-0716-1024-4
eBook Packages: Springer Protocols