| Solution Information | help | |
| Enzyme: | Estrogen receptor beta | |
| inhibitor: | BDBM42033 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | The assay is conducted in 384- well tissue culture plates with an assay volume of 20 microL. 2x10^6 Cos7 cells (ATCC #CRL-1651) are seeded in 100mm tissue culture plates and transfected with ERbeta-GFP using Lipofectamine2000 (Invitrogen; per manufacturer's instructrions). Twenty-four hours following transfection, media is changed from DMEM supplemented with 10% FBS, Pen/Strep 2 milliM L-glutamine to serum-free, phenol-red free DMEM F-12 supplemented with Pen/Strep 2 milliM L-glutamine and cells are grown an additional 24 hours. Cells are harvested via mechanical scraping and resuspended at 1x10^7/milliL in serum-free, phenol-red free DMEM F-12. The assay is conducted as follows. Column 1 contains cells with vehicle control, column 23 contains positive control (1 microM estrogen), and columns 2 and 24 contain untransfected cells. Dose-response curves are arrayed across the plate in duplicate with low concentrations in columns 3 and 14 and increasing concentrations of compounds ( | |
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