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JoVE Journal
Biochemistry

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Resin-Assisted Capture Coupled with Isobaric Tandem Mass Tag Labeling for Multiplexed Quantification of Protein Thiol Oxidation
 

Resin-Assisted Capture Coupled with Isobaric Tandem Mass Tag Labeling for Multiplexed Quantification of Protein Thiol Oxidation

Article DOI: 10.3791/62671-v 07:16 min June 21st, 2021
June 21st, 2021

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résumé

Protein thiol oxidation has significant implications under normal physiological and pathophysiological conditions. We describe the details of a quantitative redox proteomics method, which utilizes resin-assisted capture, isobaric labeling, and mass spectrometry, enabling site-specific identification and quantification of reversibly oxidized cysteine residues of proteins.

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Keywords: Protein Thiol Oxidation Resin-assisted Capture Isobaric Tandem Mass Tag Labeling Quantification Cystine Residues Sample Preparation Acetone Wash Protein Solubilization BCA Assay Centrifugal Filtration Protein Reduction DTT Buffer Exchange
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