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SILAC Based Proteomic Characterization of Exosomes from HIV-1 Infected Cells
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Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
SILAC Based Proteomic Characterization of Exosomes from HIV-1 Infected Cells

SILAC Based Proteomic Characterization of Exosomes from HIV-1 Infected Cells

DOI:
10.3791/54799-v

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10:24 min

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March 03, 2017

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, , , ,
1Brown University, 2COBRE Center for Cancer Research, Lifespan Laboratories,Rhode Island and Miriam Hospitals, 3Division of Infectious Diseases, Department of Medicine,Warren Alpert Medical School, Brown University

Capitoli

  • 00:05Titolo
  • 00:41Cell Culture and HIV-1 Infection
  • 02:11Exosome Isolation
  • 03:42Protein Extraction
  • 04:58Proteomic Data Analysis
  • 05:43Bioinformatics Verification and Characterization
  • 08:20Results: SILAC Is an Effective Method for Analyzing the Effects of HIV-1 Infection on Host Exosomal Proteomes
  • 09:08Conclusion

Summary

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Here, we describe a quantitative proteomics method using the technique of stable isotope labeling by amino acids in cell culture (SILAC) to analyze the effects of HIV-1 infection on host exosomal proteomes. This protocol can be easily adapted to cells under different stress or infection conditions.

Tags

SILACProteomic CharacterizationExosomesHIV-1 InfectionCell CultureLabelingUltracentrifugationMass Spectrometry

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