Format

Send to

Choose Destination
J Vis Exp. 2017 Jun 15;(124). doi: 10.3791/55483.

Using a Whole-mount Immunohistochemical Method to Study the Innervation of the Biliary Tract in Suncus murinus.

Author information

1
Department of Frontier Health Sciences, Graduate School of Human Health Sciences, Tokyo Metropolitan University.
2
Department of Anatomy, Tokyo Medical University.
3
Area of Regulatory Biology, Division of Life Science, Graduate School of Science and Engineering, Saitama University.
4
Department of Frontier Health Sciences, Graduate School of Human Health Sciences, Tokyo Metropolitan University; yittmniu@tmu.ac.jp.

Abstract

This work describes the whole-mount immunohistochemistry staining method in detail, using neurofilament protein antibody to label the innervation of the biliary tract in Suncus murinus (S. murinusĀ ). First, the specimen was dissected from S. murinus and fixed in 4% paraformaldehyde (PFA). Second, an enzymatic treatment and potential endogenous peroxidase inactivation were performed. The specimen was then exposed to the primary antibody, anti-neurofilament protein antibody, for 3-6 days. It was then incubated with the secondary antibody conjugated with horseradish peroxidase. The color reaction was revealed by reacting the specimen with a 3,3'-diaminobenzidine (DAB) substrate. This method can be applied to analyze the innervation of all visceral organs. Furthermore, this protocol can also be adapted to test other neuronal antibodies, but optimization of the antibodies should be done first. This method was originally introduced by Kuratani and Tanaka1,2,3.

PMID:
28654062
DOI:
10.3791/55483
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for MyJove Corporation
Loading ...
Support Center