BindingDB logo
myBDB logout

Assay Method Information

Assay Name:  GyraseB Assay
Description:  Initially, equimolar quantities of about 0.5ÁM each of E. coli gyraseA and S. aureus gyraseB were incubated for a period of 45 min with salmon sperm DNA (Sigma) in 50mM Tris (pH 7.5), 75mM ammonium acetate buffer for the reconstitution of the hybrid topoisomerases at 4 OC. Later the assay was performed in a 96-well microtiter plate as mentioned earlier (7). The assay buffer includes 50 mM Tris (pH 7.5), 75 mM ammonium acetate, 5% w/v glycerol, 0.5mM ΕDTA, 6 mM magnesiumchloride, 0.001% Triton X-100, 1 mMdithiothreitol, DNA of 2 μg/mL (~3 μM base pairs), 250ÁM ATP, 2.2nM of E. coligyrase A and S. aureusgyraseB. Reactions were performed with various drug concentration ranges for the calculation of IC50, with a negative moxifloxacin and positive novobiocin control as standards. The reaction was allowed to proceed for 60 min and was quenched by addition of 20uL of malachite green reagent (POMG-25H, Bioassay systems, USA) subsequently absorbance was read at 650nm after 20 mi
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.