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Subcellular Fractionation of Primary Chronic Lymphocytic Leukemia Cells to Monitor Nuclear/Cytoplasmic Protein Trafficking
JoVE Journal
Cancer Research
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JoVE Journal Cancer Research
Subcellular Fractionation of Primary Chronic Lymphocytic Leukemia Cells to Monitor Nuclear/Cytoplasmic Protein Trafficking

Subcellular Fractionation of Primary Chronic Lymphocytic Leukemia Cells to Monitor Nuclear/Cytoplasmic Protein Trafficking

DOI:
10.3791/60426-v

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11:39 min

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October 23, 2019

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, , ,
1Paul O’Gorman Leukaemia Research Centre, Institute of Cancer Sciences, College of Medical, Veterinary and Life Sciences,University of Glasgow

Chapters

  • 00:05Title
  • 01:22Isolation of Chronic Lymphocytic Leukemia (CLL) Cells from Patient Blood Samples
  • 03:23Preparation of Subcellular Fractions from CLL Cells
  • 05:54Preparation of Whole Cell Lysates (WCL) from CLL Cells
  • 06:32Downstream Analysis of Subcellular Fractions
  • 08:59Results: Analysis of Subcellular Fractionation of Primary Chronic Lymphocytic Leukemia Cells
  • 10:47Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol enables the optimization and subsequent efficient generation of nuclear and cytoplasmic fractions from primary chronic lymphocytic leukemia cells. These samples are used to determine protein localization as well as changes in protein trafficking that take place between the nuclear and cytoplasmic compartments upon cell stimulation and drug treatment.

Tags

Keywords: Subcellular FractionationChronic Lymphocytic LeukemiaNuclear/cytoplasmic Protein TraffickingTumor Suppressor ProteinsPathogenesisB Cell MalignanciesProtein LocalizationProtein MovementDensity Gradient CentrifugationMononuclear Cells

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