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  Label-Free Quantitative Analysis of Mitochondrial Proteomes Using the Multienzyme Digestion-Filter Aided Sample Preparation (MED-FASP) and “Total Protein Approach”

Wisniewski, J. R. (2017). Label-Free Quantitative Analysis of Mitochondrial Proteomes Using the Multienzyme Digestion-Filter Aided Sample Preparation (MED-FASP) and “Total Protein Approach”. In D. Mokranjac, & F. Perocchi (Eds.), Mitochondria (pp. 69-77). New York, NY: Humana Press.

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 Creators:
Wisniewski, Jacek R.1, Author              
Affiliations:
1Mann, Matthias / Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565159              

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Free keywords: QUANTIFICATION; DEPTHProteomic sample preparation; Multienzyme Digestion-Filter Aided Sample Preparation (MED-FASP); "Total Protein Approach"; Absolute protein quantitation; WF-assay;
 Abstract: Determination of proteome composition and measuring of changes in protein titers provide important information with a substantial value for studying mitochondria. This chapter describes a workflow for the quantitative analysis of mitochondrial proteome with a focus on sample preparation and quantitative analysis of the data. The workflow involves the multienzyme digestion-filter aided sample preparation (MED-FASP) protocol enabling efficient extraction of proteins and high rate of protein-to-peptide conversion. Consecutive protein digestion with Lys C and trypsin enables generation of peptide fractions with minimal overlap, largely increases the number of identified proteins, and extends their sequence coverage. Abundances of proteins identified by multiple peptides can be assessed by the "Total Protein Approach."

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Language(s): eng - English
 Dates: 2017
 Publication Status: Published in print
 Pages: 9
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Degree: -

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Title: Mitochondria
  Subtitle : Practical Protocols
  Other : Methods in Molecular Biology
Source Genre: Book
 Creator(s):
Mokranjac, D., Editor
Perocchi, F., Editor
Affiliations:
-
Publ. Info: New York, NY : Humana Press
Pages: - Volume / Issue: 1567 Sequence Number: - Start / End Page: 69 - 77 Identifier: ISSN: 1064-3745
ISBN: 978-1-4939-6824-4; 978-1-4939-6822-0

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Title: Methods in Molecular Biology
  Alternative Title : METHODS MOL BIOL
  Alternative Title : Methods Mol. Biol.
Source Genre: Series
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Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 1567 Sequence Number: - Start / End Page: - Identifier: ISSN: 1064-3745