System-wide immunohistochemical analysis of protein co-localization.

PloS One
Minjung KimXianjin Zhou

Abstract

The analysis of co-localized protein expression in a tissue section is often conducted with immunofluorescence histochemical staining which is typically visualized in localized regions. On the other hand, chromogenic immunohistochemical staining, in general, is not suitable for the detection of protein co-localization. Here, we developed a new protocol, based on chromogenic immunohistochemical stain, for system-wide detection of protein co-localization and differential expression. In combination with a removable chromogenic stain, an efficient antibody stripping method was developed to enable sequential immunostaining with different primary antibodies regardless of antibody's host species. Sections were scanned after each staining, and the images were superimposed together for the detection of protein co-localization and differential expression. As a proof of principle, differential expression and co-localization of glutamic acid decarboxylase67 (GAD67) and parvalbumin proteins was examined in mouse cortex. All parvalbumin-containing neurons express GAD67 protein, and GAD67-positive neurons that do not express parvalbumin were readily visualized from thousands of other neurons across mouse cortex. The method provided a global v...Continue Reading

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Citations

Feb 11, 2014·Neuropsychopharmacology : Official Publication of the American College of Neuropsychopharmacology·Victoria RisbroughXianjin Zhou
May 7, 2014·Annals of Neurology·Francesca CicchettiThomas B Freeman
Dec 21, 2012·The Journal of Comparative Neurology·James T McKennaRitchie E Brown
Apr 29, 2014·Histopathology·Michiel van den BrandJ Han J M van Krieken
Jun 4, 2015·The International Journal of Neuropsychopharmacology·Kerin K HigaXianjin Zhou
Sep 14, 2014·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Victoria L Gadd
Jul 12, 2017·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Maddalena Maria BolognesiGiorgio Cattoretti
May 12, 2020·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Maddalena Maria BolognesiGiorgio Cattoretti
May 12, 2020·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Onno J de BoerMarcel B M Teunissen
Apr 4, 2020·Journal of Neuroscience Methods·Alexander J EhrenbergLea T Grinberg

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