Search and Browse
Download
Enter Data
|
Assay Method Information |
|
| | CaMKII Cell-Based Activity |
| Description: | CaMKII activity in neonatal rat ventricular myocytes (NRVM) was measured by monitoring the phosphorylation level of phospholamban using high-content imaging. NRVMs were plated in a clear bottom 384-well plate in medium supplemented with 10% fetal bovine serum (FBS) and incubated at 37° C., 5% CO2 overnight. The medium was changed to a serum-free medium and compounds were dosed to the cells. The cells were incubated at 37° C., 5% CO2 for 4 hours and were then treated with 100 nM ATX-II (Alomone Labs) to activate CaMKII. 10 minutes after the addition of ATX-II, the cells were fixed with 4% formaldehyde followed by permeabilization with 0.1% triton X100. The fixed cells were stained with an anti-phospho-phospholamban primary antibody (Santa Cruz Biotechnology) and an Alexa488-labeled anti-rabbit IgG secondary antibody (Life Technologies Corporation) as well as Hoechst 33342 (Life Technologies Corporation). The cells were imaged using an ArrayScan VTI (Cellomics) and analyzed for cells containing a positive fluorescent signal at 485 nm, reported as % responder. The % responders were plotted against the compound concentration to determine the compounds EC50. |
| 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. |
|||||||||||||