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Small-Scale Colorimetric Assays of Intracellular Lactate and Pyruvate in the Nematode Caenorhabditis elegans
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Bioquímica
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JoVE Journal Bioquímica
Small-Scale Colorimetric Assays of Intracellular Lactate and Pyruvate in the Nematode Caenorhabditis elegans

Small-Scale Colorimetric Assays of Intracellular Lactate and Pyruvate in the Nematode Caenorhabditis elegans

DOI:
10.3791/57807-v

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09:35 min

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October 15, 2018

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, ,
1Department of Health Science, School of Sports & Health Science,Daito Bunka University, 2Department of Molecular Life Science, School of Medicine,Tokai University, 3Department of Health Management, School of Health Study,Tokai University

Capítulos

  • 00:04Título
  • 00:37Caenorhabditis elegans Synchronized Culture
  • 03:21Extraction of Cellular Fraction from C. elegans
  • 05:35Lactate, Pyruvate and Protein Assays
  • 07:32Results: Efficiency of Lactate and Pyruvate Colorimetric Assays After Protein Precipitation of C. elegans
  • 08:26Conclusion

Summary

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We describe a modified small-scale extraction and colorimetric assays of lactate and pyruvate in the nematode C. elegans. When utilizing commercial assay kits, the technical development of their sensitivity and accuracy is important. Protein precipitation in extraction is the most critical step for the quantitative determination of intracellular metabolites.

Tags

Colorimetric AssayLactatePyruvateCaenorhabditis ElegansNematodeProtein PrecipitationHomogenizationE. Coli OP50NGM Agar PlatesGravid HermaphroditesEgg IsolationL1 LarvaeSynchronous CultureColorimetric AnalysisBiochemical Methodology

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