BindingDB logo
myBDB logout

Assay Method Information

Assay Name:  In vitro HDACs Inhibition Fluorescence Assay
Description:  In brief, 10 μL of enzyme solution (HeLa nuclear extracts, HDAC1, HDAC4, HDAC6, HDAC8, HDAC11) was mixed with various concentrations of target compounds (50 μL), SAHA, and PXD101, using 100% and none HDACs groups as control group. Afterincubation at 37 C for 10 min, 40 μL of fluorogenic substrate (Boc-Lys(Ac)-AMC for HeLa nuclear extracts; Ac-Leu- GlyLyS(Ac)-AMC for HDAC1, HDAC6, and HDAC11; Ac-Leu-GlyLyS(Tfa)-AMC for HDAC4 and HDAC8) was added, and then, the mixture was incubated at 37 C for 30 min. The mixture was stopped by addition of 100 μL of developer containing trypsin and TSA afterward. Over the next incubation at 37 C for 20 min, fluorescence intensity was measured using a microplate reader at excitation and emission wavelengths of 390 and 460 nm, respectively.
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.