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Localization of Hippo Signaling Components in Drosophila by Fluorescence and Immunofluorescence

  • Cordelia Rauskolb
  • Kenneth D. IrvineEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1893)

Abstract

Visualization of in vivo protein levels and localization is essential to analysis and elucidation of Hippo signaling mechanisms and its roles in diverse tissues. This is best done by imaging proteins using fluorescent labels. Fluorescent labeling of a protein can be achieved by direct conjugation to an intrinsically fluorescent protein, like GFP, or by use of antibodies conjugated to fluorescent dyes. Immunofluorescence imaging in Drosophila typically begins with dissection and fixation of a sample tissue, followed by a series of washes and incubations with primary antibodies, directed against proteins of interest, and dye-labeled secondary antibodies, directed against the primary antibodies. This may be followed by fluorescent dyes that label cellular components, such as DNA-labeling dyes to mark nuclei. After staining and washing is completed, samples are placed in a mounting media, transferred to a microscope slide, and imaged on a confocal microscope.

Key words

Hippo Yorkie Antibody Immunofluorescence Confocal GFP 

Notes

Acknowledgments

Research in the Irvine laboratory is supported by NIH grants GM78620 and GM121537.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Waksman Institute and Department of Molecular Biology and BiochemistryRutgers UniversityPiscatawayUSA

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