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Automated Sample Multiplexing by using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)
JoVE Journal
Chemistry
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JoVE Journal Chemistry
Automated Sample Multiplexing by using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)

Automated Sample Multiplexing by using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)

DOI:
10.3791/61342-v

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09:24 min

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December 18, 2020

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1Department of Chemistry,Vanderbilt University, 2Vanderbilt Memory & Alzheimer’s Center,Vanderbilt University Medical Center, 3Vanderbilt Brain Institute,Vanderbilt University Medical Center, 4Vanderbilt Institute of Chemical Biology,Vanderbilt University Medical Center

Chapters

  • 00:04Introduction
  • 00:51Protein Extraction and Sample Preparation
  • 03:48Desalting Step 1
  • 04:39Dimethylation Labeling and Isobaric Tagging
  • 07:21Results: Automated cPILOT Processing of 22 Samples
  • 08:47Conclusion

Summary

Automatic Translation

Automatic Translation

Combined precursor isotopic labeling and isobaric tagging (cPILOT) is an enhanced sample multiplexing strategy that is capable of increasing the number of samples that can be analyzed simultaneously with available isobaric tags. Incorporation of a robotic platform has greatly increased experimental throughput, reproducibility, and quantitative accuracy.

Tags

Automated CPILOTHigh ThroughputBiological InformationAgingAlzheimer’s DiseaseLiver HomogenateUreaDenaturation BufferDTTIAMCystine

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