| Solution Information | help | |
| Enzyme: | BRCA1-associated RING domain protein 1 | |
| inhibitor: | BDBM72275 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | 1) p53-null SaOS-2 osteosarcoma cell line, expressing fusion proteins of the Brca1 (2-109 aa) and Bard1 (26-126 aa) RING domains with the N- and C- terminal fragments of the firefly luciferase from a bidirectional doxycycline-inducible promoter, was obtained from the assay provider's laboratory. 2) Saos-2 cells were expanded by passaging at 2 to 4-fold dilutions with the confluency kept between 30% - 95% in DMEM/High Modified (1x) supplemented with 10% HI FBS, 400 ug/mL G418, 10 ug/mL Cipro, 1/100 NaPyruvate, 1/100 L-Glutamine, 1/100 Penicillin/streptomycin. 3) Assay Media: Filtered DMEM/High Modified (1x) supplemented with 2% HI FBS, 400 ug/mL G418, 10 ug/mL Cipro, 1/100 NaPyruvate, 1/100 L-Glutamine, 1/100 Penicillin/streptomycin. For screening prepare Saos-2 cell suspensions at 1.67 x 105 cells/mL in fresh Assay Media 4) Steady-Glo Luciferase Assay System (Promega, Cat #E2550) 4) 1536 well white solid bottom TC treated plate (Aurora Biotechnology, Cat#00029846) Dose Response R | |
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