The typical glucose oxidase assay involves the presence of peroxidase and a chromogenic substrate (i.e. ABTS). In this way, the hydrogen peroxide produced by GOX is used by the second enzyme to convert ABTS to its oxidized form, presenting a strong absorbtion around 420 nm.

I did as Paolo Zucca mentioned and worked super. Here are the conditions:

The reaction mixture in total volume of 1 ml consisted of 0.1 M sodium phosphate pH 7.0, 5 mM 2,2´-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), 0.15 U horseradish peroxidise, 1 mM potassium hexacyanoferrate(III) (K3[Fe(CN)6]) and 10 U glucose oxidase. The oxidation of ABTS was followed at 725 nm (ε = 19 mM-1cm-1). K3[Fe(CN)6] was used to oxidize the reducing agent cysteine of the medium, which bleached the blue-green oxidized ABTS (ABTS+) (Bergmeyer and Bernt, 1974)

I usually use the coupled enzymatic assay with peroxidase and the chromogen o-dianisidine for estimating glucose oxidase activity and measure it in an spectrophotometer. This methodology is suitable also for microplates and it is a relative low cost assay. If you want the complete protocol, you can contact me by e-mail or by direct message.

The glucose oxidase enzyme (GOx) is an oxido-reductase that catalyzes the oxidation of glucose to hydrogen peroxide and D-glucono-δ-lactone. In cells, it aids in breaking the sugar down into its metabolites. The Glucose Oxidase in sample will recognize d-glucose as a specific substrate leading to proportional color development. The activity of GOx can be easily quantified colorimetrically (λ = 570 nm) or fluorometrically (Ex/Em = 535/587). This assay detects GOx activity as low as 0.01mU with our unit definition.

Hi  Ivana Djurdjevic can you send me the detailed procedure

Hi Ivana Djurdjevic

Could you send me your detailed protocol? my email is: [email protected]

Hello Carlos Diaz-Tufinio

Could you please send me the complete protocol for detection of glucose oxidase.

My e-mail id is   [email protected]

Hi  Ivana Djurdjevic can you send me the detailed procedure

How to estimate glucose oxidase activity?. Available from: https://www.researchgate.net/post/How_to_estimate_glucose_oxidase_activity [accessed Mar 24, 2015].

Hello Ivana Dejurdjevic 

could you please send me your protocol

my e-mail is [email protected]

Hello Ivana Djurdjevic, 

                                         Please also send me the detailed procedure and protocol. Will be very grateful for your help.

email: [email protected]

Hello Ivana Djurdjevic, 

                                         Could you please also send me the detailed procedure and protocol. Will be very grateful for your help.

email: [email protected]

Hello Carlos Diaz-Tufinio

Could you please send me the detailed protocol for coupled enzymatic assay for detection of glucose oxidase.

my mail-id : [email protected]

Hi Ivana Djurdjevic,  could you please send the detailed protocol for glucose oxidase assay. My email address is [email protected]. Thanks.

could you please send the detailed protocol for glucose oxidase assay. My email address is [email protected]. Thanks.