Western immunoblotting in avian shell gland sample immunoblotting methods

Department of Animal Science, University of Nebraska-Lincoln, Lincoln, Nebraska 68583, USA.

Avian shell gland tissue was subjected to Western blot analysis using anti-human estrogen receptor-alpha antibody H222. Initial attempts to obtain consistent, high-quality blots were unsuccessful because, as it turned out, excessive lipid in tissue preparations interfered with protein separation. Incremental additions of acetone eventually proved to be the critical step in solubilizing lipids and allowing consistent separation of bands on gels. A detailed description of the methodology is presented.