| Solution Information | help | |
| Enzyme: | Transcription factor p65 | |
| inhibitor: | BDBM49460 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Cell Culture: SH-5YSY (ATCC) cells stably transfected with a reporter construct containing the human NF-kB promoter fused upstream of a firefly luciferase reporter and blasticidin resistance (clone C1) were subcultured every 3-4 days in Dulbecco's Modified Eagle Medium (D-MEM) high glucose (1X) with L-glutamine and sodium pyruvate + 10% FBS + 3 ug/mL of blasticidin, and incubated at 37C in 5% CO2, and maintained at a subconfluent density of no more than 240,000 cells/cm2 for less than 30 passages. Cell Culture Reagents used: 1. D-MEM high glucose (1X), liquid, with L-glutamine and sodium pyruvate, Invitrogen: 11995-065 2. Blasticidin S HCl Invitrogen: R210-01 Compound and Control Preparation: Prior to cell addition, control drugs were diluted in assay media and added to each 384 well plate at a final concentration of 5 ng/mL for TNF-alpha (positive control). DMSO concentration of 0.3% was maintained for all control wells. Test compounds were serially diluted in a plate-to-pla | |
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