Western Blot Analysis and Immunostaining for Prediction of Embryotoxicity in Mus musculus

Methods Mol Biol. 2018:1797:497-505. doi: 10.1007/978-1-4939-7883-0_28.

Abstract

A broad range of research must be answered in order to gain a complete understanding of the histological and histochemical profile of teratological exposure in Mus musculus. Continued research is needed to track patterns of teratogen effects on the DNA expression of the embryonic brain and its variation impact. An important technique used in cell and molecular biology is Western blotting. By using a Western blot analysis and immunostaining, researchers are able to predict embryotoxicity in Mus musculus. The method uses three elements to accomplish this task: (1) Nonspecific antibody binding to a nitrocellulose membrane, (2) an incubation using a primary antibody, and (3) the antigen-antibody reaction using a secondary antibody. The proteins are further stained with a substrate/chromogen. In this chapter, the electrophoresis-based protein detection following mice embryonic exposure to a teratogen, 2-methoxyethanol, is described.

Keywords: Nitrocellulose; One-dimensional SDS gel electrophoresis; Paraffin infiltration; Rotary microtome; Western blot.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western* / methods
  • Embryo, Mammalian / drug effects*
  • Embryo, Mammalian / metabolism*
  • Immunohistochemistry* / methods
  • Mice
  • Toxicity Tests* / methods