Abstract
Automated detection of mRNAs and proteins in the same tissue sections is not a routine procedure. Successful experiment depends on the preparation of the tissue, the detection procedure, as well as the quality of the probes and antibodies. The multiplexed detections require experimental conditions, preserving the state of the molecular targets of interest and providing expression pattern of each target the same as in a single detection. Here we describe in detail the automated protocols used to detect mouse Lgr5 mRNA by in situ hybridization and immunofluorescence detection of lysozyme in the same mouse intestinal sections. Both the in situ hybridization and the protein detection were performed with an automated staining processor and provided strong and reproducible results.
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References
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Acknowledgments
We thank the members of the Molecular Cytology Core Facility for discussions and support. We would also like to thank Dr. Willie Mark from Mouse Genetics Core Facility at MSKCC for providing help and advice on mouse breeding. This work was supported by CCSG P30 CA 008748 grant from NCI.
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Turkekul, M. et al. (2017). Automated Double In Situ Detection of Mouse Lgr5 mRNA and Lysozyme Protein in Examining the Neighboring Cell Types of the Mouse Intestinal Crypt. In: Kalyuzhny, A. (eds) Signal Transduction Immunohistochemistry. Methods in Molecular Biology, vol 1554. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6759-9_19
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DOI: https://doi.org/10.1007/978-1-4939-6759-9_19
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6757-5
Online ISBN: 978-1-4939-6759-9
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