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树脂辅助捕获与同量异位串联质量标签标记相结合，用于蛋白质硫醇氧化的多重定量

JoVE Journal
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

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树脂辅助捕获与同量异位串联质量标签标记相结合，用于蛋白质硫醇氧化的多重定量

DOI:

10.3791/62671-v

•

07:16 min

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June 21, 2021

•

Matthew J. Gaffrey, Nicholas J. Day, Xiaolu Li, Wei-Jun Qian

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

Chapters

00:04Introduction
00:31Resin-Assisted Capture
03:52On-Resin Tryptic Digestion and Tandem Mass Tag (TMT) Labeling
04:45Peptide Elution
05:47Results: Analysis of Peptides from Resin-Assisted Capture Enrichment
06:35Conclusion

Summary

Automatic Translation

蛋白硫醇氧化在正常的生理和病理生理条件下具有显著意义。我们描述了定量氧化还原蛋白质组学方法的细节，该方法利用树脂辅助捕获，同量异位标记和质谱，能够对蛋白质的可逆氧化半胱氨酸残基进行位点特异性鉴定和定量。

Tags

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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