Cite as: Cold Spring Harb. Protoc.; 2008; doi:10.1101/pdb.prot5030

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High-Throughput Paraffin Section In Situ Hybridization and Dual Immunohistochemistry on Mouse Tissues

Bree Rumballe1, Kylie Georgas1, and Melissa H. Little1,2,3

1 Institute for Molecular Bioscience, University of Queensland, St. Lucia, 4072, Australia
2 Australian Stem Cell Centre, University of Queensland, St. Lucia, 4072, Australia

3Corresponding author (Melissa.Little{at}stemcellcentre.edu.au)


INTRODUCTION

Section in situ hybridization (SISH) is a high-resolution tool used to analyze gene expression patterns. This protocol utilizes the Tecan Freedom EVO150 platform to perform high-throughput SISH on paraffin sections to detect mRNA with a digoxigenin (DIG)-labeled probe. The slide is mounted and imaged before performing immunohistochemistry (IHC) on the same section. The dual reaction enables a marker of protein expression to be localized on the same section as the mRNA and facilitates more accurate annotation of the gene expression.


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