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Dynamic Proteomic and miRNA Analysis of Polysomes from Isolated Mouse Heart After Langendorff Perfusion
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Biochemistry
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JoVE Journal Biochemistry
Dynamic Proteomic and miRNA Analysis of Polysomes from Isolated Mouse Heart After Langendorff Perfusion

Dynamic Proteomic and miRNA Analysis of Polysomes from Isolated Mouse Heart After Langendorff Perfusion

DOI:
10.3791/58079-v

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07:54 min

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August 29, 2018

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, , , , ,
1The Smidt Heart Institute,Cedars-Sinai Medical Center, 2Advanced Clinical Biosystems Research Institute,Cedars-Sinai Medical Center, 3Institute of Analytical Chemistry of the Czech Academy of Sciences

Chapters

  • 00:04Title
  • 00:55Tissue Homogenization, Solubilization, and Sucrose Density Gradient Sedimentation
  • 02:49Proteomic Analysis
  • 05:36Results: Representative mRNA and Protein Distribution and Identification
  • 06:54Conclusion

Summary

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Here we present a protocol to perform polysome profiling on the isolated perfused mouse heart. We describe methods for heart perfusion, polysome profiling, and analysis of the polysome fractions with respect to mRNAs, miRNAs, and the polysome proteome.

Tags

Keywords: Dynamic ProteomicsMiRNA AnalysisPolysomesLangendorff PerfusionCardiac Stress ResponseProtein SynthesisMRNA TranslationMicroRNAProtein ExtractionSucrose GradientUltracentrifugationFraction Collection

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