Optimization of immunohistochemical detection of rat ESR2 proteins with well-validated monoclonal antibody PPZ0506.

Yujiro Hattori
Hirotaka Ishii
Shimpei Higo
Mai Otsuka
Moeko Kanaya
Keisuke Matsumoto
Mina Ozawa
Hitoshi Ozawa

Although there are few well-validated antibodies against ESR2 proteins
 a recent validation assessment identified a specific monoclonal antibody against human ESR2 proteins (PPZ0506). Furthermore
 our previous study confirmed its cross-reactivity and specificity against rodent ESR2 proteins
 enabling the determination of true ESR2 distribution profiles in rodents. Therefore
 we aimed to determine optimal conditions for ESR2 detection by PPZ0506 immunostaining and analyze ESR2 distribution in rats. We evaluated several staining conditions using paraffin-embedded and frozen ovary sections. Immunohistochemical staining with PPZ0506 antibody required strong antigen retrieval and appropriate antibody dilution. Subsequent immunohistochemical analysis in multiple tissues under optimized conditions revealed that rat ESR2 proteins are expressed in a more localized manner than previously assumed. Our results suggest that previous immunohistochemical studies using inadequately validated antibodies against ESR2 proteins overestimated their distribution profiles. We expect that optimized immunohistochemical detection with PPZ0506 antibody can help researchers solve several conflicting problems in ESR2 research.

Molecular and cellular endocrinology

523

10.1016/j.mce.2020.111145

https://doi.org/10.1016/j.mce.2020.111145
https://www.ncbi.nlm.nih.gov/pubmed/33400952