Unit

UNIT 9.28 Measurement of Cytoplasmic to Nuclear Translocation

  1. Thaddeus C. George1,
  2. Philip J. Morrissey1,
  3. Chongwei Cui2,
  4. Sukhwinder Singh2,
  5. Patricia Fitzgerald Bocarsly2

Published Online: 1 JAN 2009

DOI: 10.1002/0471142956.cy0928s47

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

George, T. C., Morrissey, P. J., Cui, C., Singh, S. and Bocarsly, P. F. 2009. Measurement of Cytoplasmic to Nuclear Translocation. Current Protocols in Cytometry. 47:9.28:9.28.1–9.28.15.

Author Information

  1. 1

    Amnis Corporation, Seattle, Washington

  2. 2

    University of Medicine and Dentistry of New Jersey, New Jersey Medical School, Newark, New Jersey

Publication History

  1. Published Online: 1 JAN 2009
  2. Published Print: JAN 2009

Abstract

A method is described for the quantitative assessment of the translocation of signaling molecules from the cytoplasm to the nucleus in cells. This method utilizes fluorochrome-conjugated antibodies to the signaling molecule and a nuclear dye, and it is based on imagery acquired rapidly in flow with the use of a multispectral imaging cytometer. The analysis correlates the spatial distribution of the stained translocating signaling molecule with nuclear staining, and it generates a quantitative score for each cell using Pearson's correlation coefficient. Examples described in this section use reagents that detect NFκB and IRF-7 and measure the translocation of these molecules under stimulating conditions. A protocol for combining cell surface phenotype with cytoplasm to nuclear translocation is also included. Curr. Protocol. Cytom. 47:9.28.1-9.28.15. © 2009 by John Wiley & Sons, Inc.

Keywords:

  • nuclear translocation;
  • transcription factors;
  • multispectral imaging