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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
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JoVE Journal Biyokimya
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

•

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

Bölümler

  • 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

Özet

Otomatik Çeviri

Otomatik Çeviri

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。

Etiketler

Protein Thiol OxidationResin-assisted CaptureIsobaric Tandem Mass Tag LabelingQuantificationCystine ResiduesSample PreparationAcetone WashProtein SolubilizationBCA AssayCentrifugal FiltrationProtein ReductionDTTBuffer Exchange

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