| Solution Information | help | |
| Enzyme: | Ubiquitin-conjugating enzyme E2 N | |
| inhibitor: | BDBM78849 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Materials: Assay buffer: 50 mM Hepes (pH 7.5), 0.1mM DTT, 0.005% Empigen, 0.1% BSA, 1.25 mM MgCl2 E1: Produced at the Sanford-Burnham Medical Research Institute's protein production core facility Fl-Ub: Invitrogen Tb-Ub: Invitrogen ATP: Sigma Ubc13: Produced at the Sanford-Burnham Medical Research Institute's protein production core facility Uev1a: Produced at the Sanford-Burnham Medical Research Institute's protein production core facility Assay plate: Corning 1536 Well White Plate (Catalogue #: 3725) I. Compound Addition: 1. Dose response curves contained 10 concentrations of compounds obtained using a twofold serial dilution. Compounds were serially diluted in 100 % DMSO (1% final DMSO concentration). Using LabCyte Echo, transfer 40 nL compounds in triplicate into assay plate columns 5 - 48. Transfer 40 nL of DMSO to positive and negative control wells in columns 1 - 4. 2. Centrifuge plates at 1000 rpm for 1 min. 3. Seal the plates and leave them at RT. No | |
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