Applicability of Anti-Human Estrogen Receptor β Antibody PPZ0506 for the Immunodetection of Rodent Estrogen Receptor β Proteins
Abstract
:1. Introduction
2. Results
2.1. Specificity and Cross-Reactivity of PPZ0506 Antibody Against Human, Mouse, and Rat ERβ Proteins
2.1.1. Construction of Expression Vectors Encoding Human, Mouse, and Rat ERβ
2.1.2. Immunoblot Analyses of Human, Mouse, and Rat ERβ in Transfected Cells
2.1.3. Immunocytochemical Analyses of Human, Mouse, and Rat ERβ in Transfected Cells
2.2. Immunohistochemical Analyses of ERβ Proteins in Rat Organs
2.2.1. Expression of ERβ Proteins in Rat Ovary
2.2.2. Expression of ERβ Proteins in Rat Prostate
2.2.3. Expression of ERβ Proteins in Rat Testis
2.2.4. Expression of ERβ Proteins in Rat Brain
2.2.5. Expression of ERβ Proteins in Rat Lung, Anterior Pituitary, Uterus and, Adrenal Gland
2.3. Alternative Promoter Usage Profiles of Human and Rat ERβ Genes
2.3.1. Genomic Organization of Human and Rat ERβ Genes
2.3.2. Alternative Promoter Usage of Human ERβ Gene
2.3.3. Alternative Promoter Usage of Rat ERβ Gene
3. Discussion
4. Materials and Methods
4.1. Animals
4.2. Total RNA Extraction, 5′-RACE, and RT-PCR
4.3. Vector Construction
4.4. Cell Culture and Transfection
4.5. Immunoblotting
4.6. Immunocytochemistry
4.7. Immunohistochemistry
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
ER | Estrogen receptor |
ERα | Estrogen receptor α |
ERβ | Estrogen receptor β |
ERE | Estrogen response element |
E2 | 17β-estradiol |
EtOH | Ethanol |
DAPI | 4,6-diamino-2-phenylindole |
AVPV | Anteroventral paraventricular nucleus |
PVH | Paraventricular nucleus of hypothalamus |
UTR | Untranslated region |
RACE | Rapid amplification of cDNA end |
ORF | Open reading frame |
FBS | Fetal bovine serum |
DMEM | Dulbecco’s modified Eagle medium |
PBS | Phosphate-buffed saline |
PB | Phosphate buffer |
TBS | Tris-buffered saline |
BSA | Bovine serum albumin |
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Ishii, H.; Otsuka, M.; Kanaya, M.; Higo, S.; Hattori, Y.; Ozawa, H. Applicability of Anti-Human Estrogen Receptor β Antibody PPZ0506 for the Immunodetection of Rodent Estrogen Receptor β Proteins. Int. J. Mol. Sci. 2019, 20, 6312. https://doi.org/10.3390/ijms20246312
Ishii H, Otsuka M, Kanaya M, Higo S, Hattori Y, Ozawa H. Applicability of Anti-Human Estrogen Receptor β Antibody PPZ0506 for the Immunodetection of Rodent Estrogen Receptor β Proteins. International Journal of Molecular Sciences. 2019; 20(24):6312. https://doi.org/10.3390/ijms20246312
Chicago/Turabian StyleIshii, Hirotaka, Mai Otsuka, Moeko Kanaya, Shimpei Higo, Yujiro Hattori, and Hitoshi Ozawa. 2019. "Applicability of Anti-Human Estrogen Receptor β Antibody PPZ0506 for the Immunodetection of Rodent Estrogen Receptor β Proteins" International Journal of Molecular Sciences 20, no. 24: 6312. https://doi.org/10.3390/ijms20246312