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Isolation and Staining of Mouse Skin Keratinocytes for Cell Cycle Specific Analysis of Cellular Protein Expression by Mass Cytometry
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Biologie du développement
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Journal JoVE Biologie du développement
Isolation and Staining of Mouse Skin Keratinocytes for Cell Cycle Specific Analysis of Cellular Protein Expression by Mass Cytometry

Isolation and Staining of Mouse Skin Keratinocytes for Cell Cycle Specific Analysis of Cellular Protein Expression by Mass Cytometry

DOI:
10.3791/59353-v

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12:34 min

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May 09, 2019

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,
1Department of Dermatology and Charles C. Gates Center for Regenerative Medicine,University of Colorado Anschutz Medical Campus

Chapitres

  • 00:04Titre
  • 01:16Preparations
  • 02:43Labeling of S phase by IdU Incorporation and Isolation of Cells
  • 05:19Cisplatin Labeling and Cell Fixation
  • 06:56Barcoding of Samples
  • 08:02Labeling of Cells for Mass Cytometry
  • 09:48Results: Cell Cycle Specific Analysis of Cellular Protein Expression by Mass Cytometry
  • 11:51Conclusion

Summary

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Traduction automatique

This protocol describes how to isolate skin keratinocytes from mouse models, to stain with metal-tagged antibodies, and to analyze stained cells by mass cytometry in order to profile the expression pattern of proteins of interest in the different cell cycle phases.

Tags

Keywords: KeratinocytesCell CycleMass CytometryProtein ExpressionCell ProliferationRecherche en cancérologieCell IsolationSkin CellsIdUParaformaldehydeCisplatin

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