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Whole-Mount In Situ Hybridization on Murine Skeletogenic Tissues

  • Timothy Rutkowsky
  • Deepika Sharma
  • Matthew J. Hilton
Part of the Methods in Molecular Biology book series (MIMB, volume 1130)

Abstract

Whole-mount in situ hybridization (WISH) is a highly sensitive method for spatial and temporal visualization of RNA transcripts throughout an entire tissue. This method is an excellent tool for studying both embryonic development and disease. It is based on sequence complementation between a Digoxigenin (DIG)-labeled antisense RNA probe and the sense RNA transcript of interest in the tissue. The labeled probe is then visualized by immunohistochemical methods using an antibody conjugated to alkaline phosphatase that catalyzes a color reaction.

Keywords

Whole-mount in situ hybridization Digoxigenin RNA Gene expression 

Notes

Acknowledgements

This work was in part supported by the following NIH grants: R01 grants (AR057022 and AR063071), R21 grant (AR059733), T32 training grant (AR053459), and P30 Core Center grant (AR061307).

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Copyright information

© Springer Science+Business Media, LLC 2014

Authors and Affiliations

  • Timothy Rutkowsky
    • 1
  • Deepika Sharma
    • 1
  • Matthew J. Hilton
    • 1
  1. 1.University of Rochester Medical CenterRochesterUSA

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