Whole-Mount In Situ Hybridization (WISH) for Salamander Embryos and Larvae

Sruthi Purushothaman, Ashley W. Seifert

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Scopus citations


Whole-mount in situ hybridization (WISH) is widely used to visualize transcribed gene sequences (mRNA) in developing embryos, larvae, and other nucleotide probe permeable tissue samples. This methodology involves the hybridization of an antisense nucleotide probe to the target mRNA, followed by chromogen or fluorescence-based detection. Here we describe a protocol for the spatiotemporal analysis of mRNA transcripts in axolotl embryos/larvae using digoxigenin-labeled riboprobes, anti-digoxigenin alkaline phosphatase, Fab fragments antibody, and NBT/BCIP chromogen detection.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
Number of pages13
StatePublished - 2023

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Bibliographical note

Publisher Copyright:
© 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.


  • Digoxigenin
  • In situ hybridization (ISH)
  • In vitro transcription
  • Limb development
  • Limb regeneration
  • Riboprobe
  • Salamander

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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