Blocking Cross-Species Secondary Binding When Performing Double Immunostaining With Mouse and Rat Primary Antibodies.

Frontiers in Neuroscience
Shanping MaoAkiva S Cohen

Abstract

Immunostaining is a powerful technique and widely used to identify molecules in tissues and cells, although critical steps are necessary to block cross-reaction. Here we focused on an overlooked cross immunoreactivity issue where a secondary antibody (secondary) cross-reacts with a primary antibody (primary) from a different species. We first confirmed the previously reported cross-species binding of goat anti-mouse secondary to rat primary. This was accomplished by staining with a rat primary against glial fibrillary acidic protein (GFAP) and visualizing with goat (or donkey) anti-mouse secondary. We then further revealed the converse cross-species binding by staining with a mouse primary against neuronal nuclear protein (NeuN) and visualizing with anti-rat secondaries. We speculate that mouse and rat primaries share antigenicity, enabling either secondary to recognize either primary. To block this cross-species binding in double staining experiments, we compared three protocols using mouse anti-NeuN and rat anti-GFAP, two primaries whose antigens have non-overlapping distributions in brain tissues. Simultaneous staining resulted in cross-species astrocytic staining (anti-mouse secondary to rat anti-GFAP primary) but no cross-...Continue Reading

References

Jan 1, 1993·Methods in Cell Biology·P A EricksonS K Fisher
Mar 30, 2007·Molecular and Cellular Neurosciences·Jill SeeJudith B Grinspan
Sep 12, 2008·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Andrea Lorincz, Zoltan Nusser
Oct 16, 2008·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Clifford B Saper
Nov 1, 2007·Blood Transfusion = Trasfusione Del Sangue·Roberto Reverberi, Lorenzo Reverberi
Nov 21, 2014·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Guoxiang XiongNoam A Cohen
Dec 1, 2015·Frontiers in Neurology·Colin J SmithAkiva S Cohen

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