High Density Receptor Ligand Binding Assays in the MultiScreen®HTS 384-well Glass Fiber Filter Plate
Poster Jun 07, 2005
Gertrud Beterams, Steven Sheridan, Sonia Gil and Libby Kellard
Radiometric filter binding assays are acknowledged for their ability to identify and confirm lead compounds in drug discovery.They can be limited in their utility due to throughput, cost, andamenability to automation. Data obtained using a new 384 well filter plate demonstrate that automated quantitative and screening assays can be developed that are equivalent to assays run in a 96-well format.
Using the Muscarinic M1 G-protein Coupled Receptor as a model system, we achieved accurate and reproducible determinations of binding affinity (Kd) and IC50 results for known ligands on the 384-well plate device.
As a result of being able to incubate the reaction mixture in the filter plate and configure the assay using half the reaction volume, significant reductions in reagent costs and radioactive waste were achieved. Use of the fully-automated 384 filter plate makes it possible to develop higher throughput, lower cost radiometric receptor binding assays.
We utilized paired synthetic crRNAs coupled with our synthetic tracrRNA in cells transduced with lentiviral Cas9 to perform a functional knockout on hsa-miR-221. This three-part system (crRNA, tracrRNA and Cas9) has demonstrated efficient gene editing when used with only one guide RNA, but the goal was to use two crRNAs to remove the entire stem-loop.READ MORE