Abstract
This text provides a brief overview of specific issues in the meat industry which may be resolved using the current proteomics-based methodology. Proteins as key to understanding biological processes as well as to change the characteristics of meat products offer enormous potential in the context of meat science. The quantification, analysis and curation of proteomics data, and particularly, the industrial applications of these new findings will become a more essential tool in the coming years.
The methodology described in this chapter presents the basis for quantification, analysis, and curation of proteomics data, representing the pillars of main proteomic methods. The main steps include 1. Extraction and quantification of proteins; 2. Separation of proteins by isoelectric point; 3. Separation of proteins by molecular weight; 4. Staining of proteins; 5. Tryptic digestion with enzymes; and 6. identification of proteins by mass spectrometry.
More recent proteomics approaches are beyond the scope of this chapter. Therefore, all steps are described in detail so that the conditions proposed by us can be reproduced by other researcher groups. Although protocols and methodology are specifically obtained from our lab, these can be easily adapted for every laboratory.
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Abbreviations
- 2-DE :
-
Two-dimensional electrophoresis
- CHAPS detergent:
-
(3-((3-cholamidopropyl) dimethylammonium)-1-propanesulfonate)
- DTT:
-
Dithiothreitol
- IPG:
-
Immobilized pH gradient
- MALDI-TOF :
-
Matrix-assisted laser/desorption ionization-time of flight
- Mr.:
-
Molecular weight
- pI:
-
Isoelectric point
- PMF:
-
Peptide Mass Fingerprint
- PSA:
-
Persulfate ammonium
- SDS:
-
Sodium dodecyl sulfate
- SDS-PAGE :
-
Sodium dodecyl sulfate polyacrylamide gel electrophoresis
- TCA:
-
Trichloroacetic
- TEMED:
-
Tetramethylethylenediamine
- Tris:
-
Tris(hydroxymethyl)aminomethane
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López-Pedrouso, M., Lorenzo, J.M., Franco, D. (2022). Proteomics. In: Lorenzo, J.M., Domínguez, R., Pateiro, M., Munekata, P.E. (eds) Methods to Assess the Quality of Meat Products. Methods and Protocols in Food Science . Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2002-1_13
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DOI: https://doi.org/10.1007/978-1-0716-2002-1_13
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