Search and Browse
Download
Enter Data
|
Assay Method Information |
|
| | Radioligand Binding Assay |
| Description: | Aliquots of radioligand are dispensed into the wells of 96-well plates. Then, duplicate aliquots of the test and reference compound dilutions are added. Finally, crude membrane fractions of cells expressing recombinant target are dispensed into each well. The reactions are incubated at room temperature and shielded from light (to prevent photolysis of light-sensitive ligands) for 1.5 hours, then harvested by rapid filtration onto Whatman GF/B glass fiber filters using a 96-well Brandel harverster. Four rapid washes are performed with chilled Standard Binding Buffer to reduce non-specific binding. Filters are placed in scintillation tubes, and counted by liquid scintillation counting. Binding data were analyzed using Prism (GraphPad, Dan Diego, CA). The log IC50 (i.e., the log of the ligand concentration that reduces radioligand binding by 50%) is estimated from the data and used to obtain the Ki by applying the Cheng-Prusoff approximation. |
| Affinity data for this assay | |
|---|---|
| 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. |
|||||||||||||