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Cell Fractionation of U937 Cells by Isopycnic Density Gradient Purification
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JoVE Journal Bioquímica
Cell Fractionation of U937 Cells by Isopycnic Density Gradient Purification

Cell Fractionation of U937 Cells by Isopycnic Density Gradient Purification

DOI:
10.3791/62119-v

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10:19 min

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August 12, 2021

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

Capítulos

  • 00:04Introduction
  • 00:37Cytosolic Protein Isolation
  • 02:45Cell Homogenization
  • 03:55Debris Removal and Crude Mitochondrial and Membrane Fraction Isolation
  • 04:47Isopycnic Density Gradient Purification
  • 06:34Nuclear Protein Isolation
  • 07:57Results: Fractionation of U937 Cells by Isopycnic Density Gradient Purification
  • 09:34Conclusion

Summary

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This fractionation protocol will allow researchers to isolate cytoplasmic, nuclear, mitochondrial, and membrane proteins from mammalian cells. The latter two subcellular fractions are further purified via isopycnic density gradient.

Tags

Keywords: Cell FractionationU937 CellsIsopycnic Density Gradient PurificationCytoplasmNucleusMitochondriaMembraneCytosolic Protein IsolationNuclear Protein ExtractionCellular CompartmentsCell LysisCell HomogenizationDensity Gradient Centrifugation

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