Good Old-Fashioned Protein Concentration Determination by Amino Acid Analysis.

Acid hydrolysis Amino acid analysis Protein concentration determination Quantitative proteome analysis

Journal

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969

Informations de publication

Date de publication:
2021
Historique:
entrez: 5 5 2021
pubmed: 6 5 2021
medline: 24 6 2021
Statut: ppublish

Résumé

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.

Identifiants

pubmed: 33950480
doi: 10.1007/978-1-0716-1024-4_2
doi:

Substances chimiques

Amino Acids 0
Proteins 0
Proteome 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

21-28

Références

Lowry OH, Rosebrough NJ, Farr AL et al (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193(1):265–275
doi: 10.1016/S0021-9258(19)52451-6
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: 10.1016/0003-2697(76)90527-3
Gornall AG, Bardawall CJ, David MM (1949) Determination of serum proteins by means of the biuret reaction. J Biol Chem 177(2):751–766
doi: 10.1016/S0021-9258(18)57021-6
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
doi: 10.1016/j.dib.2019.103742

Auteurs

Caroline May (C)

Medizinisches Proteom-Center (MPC), Medical Faculty, Ruhr-University Bochum, Bochum, Germany.
Medical Proteome Analysis, Center for Proteindiagnostics (PRODI), Ruhr-University Bochum, Bochum, Germany.

Bettina Serschnitzki (B)

Medizinisches Proteom-Center (MPC), Medical Faculty, Ruhr-University Bochum, Bochum, Germany.
Medical Proteome Analysis, Center for Proteindiagnostics (PRODI), Ruhr-University Bochum, Bochum, Germany.

Katrin Marcus (K)

Medizinisches Proteom-Center (MPC), Medical Faculty, Ruhr-University Bochum, Bochum, Germany. katrin.marcus@rub.de.
Medical Proteome Analysis, Center for Proteindiagnostics (PRODI), Ruhr-University Bochum, Bochum, Germany. katrin.marcus@rub.de.

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Classifications MeSH