Insufficient antibody validation challenges oestrogen receptor beta research.
Sandra AnderssonMårten SundbergNusa PristovsekAhmed IbrahimPhilip JonssonBorbala KatonaCarl-Magnus ClaussonAgata ZiebaMargareta RamströmOla SöderbergCecilia WilliamsAnna AsplundPublished in: Nature communications (2017)
The discovery of oestrogen receptor β (ERβ/ESR2) was a landmark discovery. Its reported expression and homology with breast cancer pharmacological target ERα (ESR1) raised hopes for improved endocrine therapies. After 20 years of intense research, this has not materialized. We here perform a rigorous validation of 13 anti-ERβ antibodies, using well-characterized controls and a panel of validation methods. We conclude that only one antibody, the rarely used monoclonal PPZ0506, specifically targets ERβ in immunohistochemistry. Applying this antibody for protein expression profiling in 44 normal and 21 malignant human tissues, we detect ERβ protein in testis, ovary, lymphoid cells, granulosa cell tumours, and a subset of malignant melanoma and thyroid cancers. We do not find evidence of expression in normal or cancerous human breast. This expression pattern aligns well with RNA-seq data, but contradicts a multitude of studies. Our study highlights how inadequately validated antibodies can lead an exciting field astray.
Keyphrases
- estrogen receptor
- poor prognosis
- rna seq
- binding protein
- single cell
- endoplasmic reticulum
- endothelial cells
- breast cancer cells
- small molecule
- induced apoptosis
- high throughput
- long non coding rna
- protein protein
- gene expression
- induced pluripotent stem cells
- stem cells
- cell proliferation
- cell therapy
- metabolic syndrome
- adipose tissue
- oxidative stress
- signaling pathway
- big data
- pi k akt
- multiple myeloma
- mesenchymal stem cells
- artificial intelligence
- cell cycle arrest
- data analysis