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TMT Sample Preparation for Proteomics Facility Submission and Subsequent Data Analysis
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
TMT Sample Preparation for Proteomics Facility Submission and Subsequent Data Analysis

TMT Sample Preparation for Proteomics Facility Submission and Subsequent Data Analysis

DOI:
10.3791/60970-v

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07:44 min

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June 08, 2020

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, , ,
1Department of Pediatrics,University of Florida

Chapters

  • 00:04Introduction
  • 00:31Protein Extraction
  • 01:40Reducing/Alkylating Reagent Treatment
  • 02:39Methanol/Chloroform Precipitation
  • 04:20Protein Digestion
  • 04:56Peptide Labeling
  • 05:57Results: Representative Protein Quantification and Volcano Plot and Protein Analysis through Evolutionary Relationships (PANTHER) Analyses
  • 07:04Conclusion

Summary

Automatic Translation

Automatic Translation

We present an optimized tandem mass tag (TMT) labeling protocol that includes detailed information for each of the following steps: protein extraction, quantification, precipitation, digestion, labeling, submission to a proteomics facility, and data analyses.

Tags

Keywords: TMT Sample PreparationProteomicsProtein ExtractionLabeling EfficiencyReproducibilityData AcquisitionCHAPS Lysis BufferBead DisruptorSonicationReducing/alkylating Reagent TreatmentTCEPIodoacetamideMethanol/chloroform Precipitation

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