BindingDB logo
myBDB logout

Assay Method Information

Assay Name:  HTRF kinase assay
Description:  The c-Met kinase activity was evaluated by homogeneous time-resolved fluorescence (HTRF) assays. In a 384-well plate, 20 µg/mL (Glue, Tyr) 4:1 was added, followed by the addition of 50 µL of 10 mM ATP solution diluted in kinase reaction buffer (50 mM HEPES, pH 7.0, 1 M DTT, 1 M MgCl2, 1 M MnCl2, and 0.1% NaN3) into each well. Various concentrations of compounds diluted in 10 µL of 1% DMSO (v/v) are used as the negative control. The reaction was initiated by the addition of purified c-Met kinase diluated in 39 µL of kinase reaction buffer solution. The reaction was incubated for 30 min at 25 °C and stopped by the addition of 5 µL of Streptavidin-XL665 and 5 µL Tk Antibody Cryptate working solution to all of the wells. The plate was read at 320 nm and 615 nm.
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.