new BindingDB logo
myBDB logout

Assay Method Information

Assay Name:  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

 
Last update November 1, 2007
©2000 BindingDB. All rights reserved.