title: Measurement of cytoplasmic to nuclear translocation.
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 NFkappaB 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.
authors.affiliation: Amnis Corporation, Seattle, Washington.
authors.forename: Thaddeus C
authors.initials: TC
authors.lastname: George
authors.affiliation: Amnis Corporation, Seattle, Washington.
authors.forename: Philip J
authors.initials: PJ
authors.lastname: Morrissey
authors.affiliation: University of Medicine and Dentistry of New Jersey, New Jersey Medical School, Newark, New Jersey.
authors.forename: Chongwei
authors.initials: C
authors.lastname: Cui
authors.affiliation: University of Medicine and Dentistry of New Jersey, New Jersey Medical School, Newark, New Jersey.
authors.forename: Sukhwinder
authors.initials: S
authors.lastname: Singh
authors.affiliation: University of Medicine and Dentistry of New Jersey, New Jersey Medical School, Newark, New Jersey.
authors.forename: Patricia Fitzgerald
authors.initials: PF
authors.lastname: Bocarsly
mesh_terms: D021581:Active Transport, Cell Nucleus; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D012107:Research Design
chemical_list: D005456:Fluorescent Dyes; D047908:Intracellular Signaling Peptides and Proteins