Search and Browse
Download
Enter Data
| Solution Information | help | |
| Enzyme: | Large T antigen | |
| inhibitor: | BDBM36880 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | The confirmatory assay was conducted in 384-well black, clear-bottom microplates. Assay buffer was 100mM Tris-HCl, 20mM KCl, 6mM MgCl2, and 0.1mg/mL BSA, pH 7.4. Initially, 4ul of test compound (20uM final concentration) in 4% DMSO/assay buffer was added to the respective plate wells. Total reaction volume (without endpoint reagent) was 20ul. Total read volume (with endpoint reagent) was 80 ul. Purified TAg was diluted to a concentration of 0.0144 ug/ul in assay buffer. 12ul of TAg (14.4ng/ul) in assay buffer was added to each well and allowed to incubate for 30m at ambient temperature. Following incubation, 5ul of 5mM ATP (A7699, Sigma-Aldrich, St. Louis, MO) in assay buffer was added to the respective wells, and incubated for 120m at ambient temperature. Final DMSO concentration was 1.0%. Endpoint addition and read: After incubation, the ADPHunter-Plus Kit (DiscoverX, Freemont, CA) was used to determine the extent of TAg-induced ATP hydrolysis. 20ul of Reagent A, immediately foll | |
| If you find an error in this entry please send us an E-mail | ||
|
Home |
| |
Search |
| |
Deposit |
| |
SiteMap |
| |
About us |
| |
Email us |
| |
Info |
|
©2000 BindingDB. All rights reserved. |
|||||||||||||