Immunofluorescent Visualization of BMP Signaling Activation on Paraffin-Embedded Tissue Sections

Maaike Alkema; Marie-José Goumans; Boudewijn P T Kruithof

doi:10.1007/978-1-4939-8904-1_14

Immunofluorescent Visualization of BMP Signaling Activation on Paraffin-Embedded Tissue Sections

Methods Mol Biol. 2019:1891:191-200. doi: 10.1007/978-1-4939-8904-1_14.

Authors

Maaike Alkema¹, Marie-José Goumans¹, Boudewijn P T Kruithof^{2

3

4}

Affiliations

¹ Department of Cell and Chemical Biology, Leiden University Medical Center, Leiden, The Netherlands.
² Department of Cell and Chemical Biology, Leiden University Medical Center, Leiden, The Netherlands. B.P.T.Kruithof@lumc.nl.
³ Department of Cardiology, Leiden University Medical Center, Leiden, The Netherlands. B.P.T.Kruithof@lumc.nl.
⁴ Netherlands Heart Institute, Utrecht, The Netherlands. B.P.T.Kruithof@lumc.nl.

PMID: 30414134
DOI: 10.1007/978-1-4939-8904-1_14

Abstract

Immunohistochemistry allows the detection of the presence, localization, and activation of proteins in biological tissues by using the ability of antibodies to bind to specific antigens. Cellular signaling can be visualized using antibodies raised against phosphorylated proteins. Phosphorylated Smad1, Smad5, and Smad9 are the activated signaling molecules of the BMP pathway that transfer BMP signals from the cell surface to the nucleus. Here we describe the detection of phospho-Smad1/5/9 on paraformaldehyde-fixed and paraffin-embedded tissue sections by immunofluorescence.

Keywords: BMP signaling; Immunohistochemistry; Phospho-Smad1; Phospho-Smad5; Phospho-Smad8/9; Tissue sections.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bone Morphogenetic Proteins / metabolism
Fluorescent Antibody Technique*
Immunohistochemistry*
Signal Transduction*

Substances

Bone Morphogenetic Proteins