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Density Gradient Ultracentrifugation for Investigating Endocytic Recycling in Mammalian Cells
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生物化学
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JoVE 杂志 生物化学
Density Gradient Ultracentrifugation for Investigating Endocytic Recycling in Mammalian Cells

Density Gradient Ultracentrifugation for Investigating Endocytic Recycling in Mammalian Cells

DOI:
10.3791/62621-v

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05:13 min

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June 30, 2021

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, ,
1School of Life Sciences,The Chinese University of Hong Kong

Chapters

  • 00:05Introduction
  • 00:33Mammalian Cell Culture, Transfection and Cell Harvest
  • 02:07Density Gradient Column Preparation
  • 02:46Fractionation and Recovery of Recycling Endosome-Containing Fraction
  • 03:17Results: Density Gradient Ultracentrifugation for Investigating Endocytic Recycling in Mammalian Cells
  • 04:47Conclusion

Summary

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This paper aims to present a protocol for preparing recycling endosomes from mammalian cells using sucrose density gradient ultracentrifugation.

Tags

Density Gradient UltracentrifugationEndocytic RecyclingMammalian CellsProtein TraffickingRecycling EndosomesCell FractionationSucrose GradientPost-nuclear SupernatantHomogenization BufferDounce HomogenizerProtease InhibitorPhosphatase Inhibitor

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