Search and Browse
Download
Enter Data
|
Assay Method Information |
|
| | EGFR IC50 Determination |
| Description: | The kinase reaction for EGFR consisted of BSA-supplemented kinase buffer, kinase, peptide, and ATP. For IC50 determinations, 10 different concentrations (ranging from 0.1 nM to 10 uM) of inhibitor were used in duplicate. Each experiment was repeated at least twice. Following the kinase reaction, Development Solution (included with the kit) was added to cleave the remaining unphosphorylated peptide. Following a 1 h incubation time with Development Solution, Stop Solution (included with the kit) was added to the reaction mixture. Fluorescence was then measured with a Spectramax M5 plate reader from Molecular Devices or a Safire2 from Tecan. Upon excitation of coumarin at 400 nm, fluorescence emission was measured at 445 nm (coumarin) and 520 nm (fluorescein). The ratio of coumarin to fluorescein emission was used to calculate the percentage of phosphorylation of the peptide by the kinase. |
| 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. |
|||||||||||||