Details and optimised conditions for MED12, ERα and ERβ antibodies
| Antibody | Source | Clone | Dilution | Incubation time | Pretreatment | Automation system | Localisation | Threshold defined |
|---|---|---|---|---|---|---|---|---|
| MED12 | Bethyl Laboratories (A300-774A) | Polyclonal | 1:500 | 30 min | Citrate pH 6.0 at 120°C for 5 min | Dako Autostainer Plus | Nucleus | High: H-score 150 and above |
| ERα | Neomarker (NM RM9101-S) | SP1 | 1:100 | 32 min | Mild CC1 pH 9.0 at 100°C for 30 min | Ventana BenchMark ULTRA | Nucleus | Positive: at least 1% immunoreactive tumour nuclei |
| ERβ | Abcam (ab288) | 14C8 | 1:200 | 1 h | Citrate pH 6.0 at 98°C for 15 min | Dako Autostainer Plus | Nucleus and cytoplasm | Positive: at least 1% immunoreactive tumour nuclei |
ER, oestrogen receptor.