Double in situ detection of sonic hedgehog mRNA and pMAPK protein in examining the cell proliferation signaling pathway in mouse embryo Journal Article

  


Authors: Fujisawa, S.; Turkekul, M.; Barlas, A.; Fan, N.; Manova, K.
Article Title: Double in situ detection of sonic hedgehog mRNA and pMAPK protein in examining the cell proliferation signaling pathway in mouse embryo
Abstract: Double in situ detection of RNA molecules and proteins in tissue sections is not trivial. A successful experiment heavily depends on the preparation of the tissue as well as the quality of the probes and antibodies. Detection of two or more molecular markers also requires reagents and experimental conditions that will preserve authenticity (accuracy) of the single staining patterns. Here, we describe in detail the protocols used to detect sonic hedgehog (Shh) mRNA by in situ hybridization and immunofluorescence staining for phosphorylated mitogen-activated protein kinase (pMAPK) in the same mouse embryonic tissue sections. In addition to protocols for manual immuno-staining, we provide data from automated machine-based staining protocols and highly recommend it to achieve strong signal and reproducible results.
Keywords: signal transduction; mitogen activated protein kinase; genetics; methodology; cell proliferation; mouse; animal; metabolism; animals; mice; in situ hybridization, fluorescence; sonic hedgehog protein; hedgehog proteins; animal embryo; fluorescent antibody technique; fluorescence in situ hybridization; messenger rna; rna, messenger; embryo, mammalian; mitogen-activated protein kinases; ultrastructure; shh protein, mouse
Journal Title: Methods in Molecular Biology
Volume: 717
ISSN: 1064-3745
Publisher: Humana Press Inc  
Date Published: 2011-01-01
Start Page: 257
End Page: 276
Language: English
PUBMED: 21370036
PROVIDER: scopus
DOI: 10.1007/978-1-61779-024-9_15
DOI/URL:

http://www.scopus.com/inward/record.url?eid=2-s2.0-84855510230&partnerID=40&md5=cfbf82d61eff4bdb50f2f89e081f6de6
Notes: Chapter 15 in "Signal Transduction Immunohistochemistry: Methods and Protocols" (ISBN: 978-1-61779-023-2) --- Export Date: 1 March 2012 - Source: Scopus
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MSK Authors
  1. Afsar Barlas
    35 Barlas
  2. Katia O Manova-Todorova
    161 Manova-Todorova
  3. Sho Fujisawa
    32 Fujisawa
  4. Mesruh Turkekul
    19 Turkekul
  5. Ning Fan
    18 Fan
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