Trypanosoma brucei Project
6/25/2005
6/25/2005
10/10/2005
This data is from a screen of 3118 kinase inhibitors from the ChemDiv library screened at 1 µM against T. brucei blood stream forms in a HTP format. Compounds with greater than 40 % inhibition of cell growth at 44 hours are listed.
Protocol
Adaptation of live parasite assay to microtiter plate format for higher throughput
The blood stream form of T. brucei parasites, growing in mid-log phase are diluted to 1 x 105 cells/ml in HMI-9 medium and aliquoted into 96 well culture plates. Each well contains inhibitor at 1 micromolar in DMSO (final concentration in assay < 1%), in an equal volume of culture medium. Control wells contain DMSO, a solvent control, or 17-DMAG (a geldanomycin derivative), a trypanocidal drug control. Parasites were incubated for ~ 44 hours at 37°C with 5% CO2. Parasite viability is determined by measuring ATP content using CellTiter-Glo™ (Promega) in a luminometer. Percent inhibition is calculated in comparison with DMSO controls.