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Cell Fractionation of U937 Cells in the Absence of High-speed Centrifugation
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Biochemistry
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JoVE Journal Biochemistry
Cell Fractionation of U937 Cells in the Absence of High-speed Centrifugation

Cell Fractionation of U937 Cells in the Absence of High-speed Centrifugation

DOI:
10.3791/59022-v

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06:53 min

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January 18, 2019

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, , , , ,
1Department of Basic and Clinical Sciences,Albany College of Pharmacy and Health Sciences

Chapters

  • 00:04Title
  • 00:35PBS Wash
  • 01:01Cytosolic Protein Isolation
  • 02:21Cell Homogenization
  • 03:32Differential Centrifugation
  • 04:58Results: Undifferentiated U937 Cells Successfully Fractionated
  • 06:15Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we present a protocol to isolate the plasma membrane, cytoplasm and mitochondria of U937 cells without the use of high-speed centrifugation. This technique can be used to purify subcellular fractions for subsequent examination of protein localization via immunoblotting.

Tags

Cell FractionationU937 CellsHigh-speed CentrifugationProtocolSubcellular FractionsCytoplasmMitochondriaPlasma MembraneHypotonic BuffersDigitoninMechanical LysisDifferential CentrifugationProtein LocalizationInexpensiveStraightforward

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