| Solution Information | help | |
| Enzyme: | X-box-binding protein 1 | |
| inhibitor: | BDBM67751 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | The top 2091 positive compound identified from HTS of small molecule inhibitors of CHOP to regulate the unfolded protein response to ER stress (AID 2732), were tested in the confirmatory assay (triplicates at 10 uM in the primary assay). The 940 positives (>70% inhibition) from the confirmatory assay were further tested in dose-response using counterscreen assay (CHO-XBP1 cells). The dose-response tests were carried out at 7 concentrations and each concentration was tested in triplicates. Cell lines & Reagents: 1. Cell line for counter screen assay (dose-response): CHO cells are transfected with a reporter that contains the 26bp intron from XBP1 placed into the 5' end of luciferase (provided by assay PI). Luciferase is not translated unless the intron is removed via IRE1=mediated splicing upon activation of the UPR. 2. Cell growth media (Ham's F12 + Glutamax, 10% FBS, 1X non-essential amino acids, and penicillin/streptomycin) (Invitrogen) 3. Tunicamycin (Calbiochem) 4. Stea | |
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