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Biochemistry

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JoVE Journal Biochemistry
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 •
1, 1, 2, 1
1Integrative Omics, Biological Sciences Division, Pacific Northwest National Laboratory, 2Bioproducts Sciences and Engineering Laboratory, Department of Biological Systems Engineering, Washington State University

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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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Resin-assisted Capture Isobaric Tandem Mass Tag Labeling Protein Thiol Oxidation Oxidized Cystine Residues Quantitative Analysis Site-specific Identification Acetone Wash Micro Pipette Glass Serological Pipette Buffer B Sonication Bicinchoninic Acid Assay Protein Concentration Centrifugal Filter
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