Search and Browse
Download
Enter Data
|
Assay Method Information |
|
| | Human Adenosine A1 Receptor Binding Assay |
| Description: | Test compound is weighed, dissolved in DMSO to make a stock solution of 10 mM, diluted with DMSO to prepare working solutions, then 100-fold diluted to the indicated concentrations. Human recombinant adenosine A1 receptors expressed in CHO-K1 cells were used to prepare membranes in incubation buffer (20 mM HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) pH 7.4, 10 mM MgCl2, 100 mM NaCl). Test compound or vehicle (2.2 μL) was added in 200 μL of membranes and incubated with 20 μL of 1 nM [3H] DPCPX (8-cyclopentyl-1,3-dipropylxanthine) for 90 minutes at 25 C. Non-specific binding was estimated in the presence of 100 μM R(−)-PIA ((R) N-(1-methyl-2-phenylethyl)adenosine). The incubation was stopped by vacuum filtration onto 0.3% PEI (polyethylenimine) presoaked GF/B filters using a harvester followed by four washes with ice-cold 50 mM Tris-HCl, pH 7.4, and the radioactivity on GF/B filtermats counted in a scintillation counter (PerkinElmer Topcount ) to determine [3H] DPCPX specifically bound. Final concentration of vehicle DMSO is 1%. |
| 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. |
|||||||||||||