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Heterokaryon Technique for Analysis of Cell Type-specific Localization

16.3K Views

09:31 min

March 11th, 2011

March 11th, 2011

16,289 Views

0:05

Title

1:00

Introduction

1:34

Cell Transfection - Method 1: Nucleofection

3:34

Cell Transfection - Method 2: Effectene

5:43

Cell Fusion

6:49

Fluorescence Microscopy

7:44

Analysis of Cell-type Specific Protein Localization

8:48

Conclusion

A flexible and efficient method for the characterization of cell type-specific protein localization and nucleocytoplasmic shuttling is described. This heterokaryon approach uses fluorescently-labeled fusion proteins to image protein localizations after cell fusion. The protocol is amenable to steady-state localizations or more dynamic determinations based on live cell imaging.

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