Superoxide Dismutase (SOD) Activity Assay Kit: Measure SOD Activity in Tissues, Cells, Blood etc.Superoxide dismutase (SOD) is one of the most important antioxidative enzymes. It catalyzes the dismutation of the superoxide anion into hydrogen peroxide and molecular oxygen. A graphical representation of data will show percent inhibition versus SOD activity U/ml. If you are measuring the reactants after complete reaction then more inhibition means lesser optical density, if it is overshooting and becoming higher than it means inhibition is going down.

Well, As far as I am familiar with the SOD Assay using NBT, It involves the production of Blue Formazan in the presence of light. The enzyme SOD breaks this complex resulting in lower OD values. Therefore, Lower values will indicate higher SOD activity and subsequent enzyme.  

Secondly, You need to prepare two blanks at a time, both without enzyme extract. One should be placed in dark and will be then used for Auto zero calibration. The second blank will represent the maximum amount of blue formazan complex formed (Indicating zero SOD activity) which will be highest value and it will be used to calculate enzyme activity. Moreover, Prepare second blank with every batch of SOD assay.

Lastly, Superoxide radicals are generated in PMS-NADH systems by oxidation of NADH and assayed by the reduction of nitroblue tetrazolium (NBT). The reduction of NBT actually results in the formation of colored complex (Blue Formazan) which is actually inhibited by SOD.

Hope this explains your Questions

Hi,

instead of more or less expensive commercially availabe kits, why don`t you try the method described by Flohè/Ötting. This method works well and is much cheaper.

-> PubMed-ID: 6328209

Thank u all for all your valuable answers

Generally, my answers resort to bibliographic references; thus, for your question I suggest these two articles and I hope to help you:

http://www.incda-fundulea.ro/rar/nr30/rar30.21.pdf

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2830880/pdf/nihms173323.pdf

Best wishes!

You can try Ingel assay. You have to run sample along with control in a NATIVE PAGE. You can easily find out the activity by measuring the achromatic zone in your gel.

 Hi,

i am doing sod assay in animal tissue, i use the protocol NADH/ NBT method, PMS+NBT+ NADH  incubates at 30 degree celsius for 90 seconds, the blue colour formed after the incubation i added glacial acetic acid and n- butanol, the color is disappeared.

my question is 

1.why the color was disappeared?

2. which rpm  to use for centrifuge?

.please tell ur answer

 thank u