I am performing the SOD assay using the pyrogallol protocol by Marklund and Marklund for estimating SOD from serum. I am not getting the desired results. I'm using a 30 mM concentration of pyrogallol, but my reading of serum is more than the pyrogallol reading.
Can anyone suggest any solutions.
i think so from my point of view timing of auto oxidation of pyrogallol is very important , like 0-1, 1-2. 2-3 sec , you have to take OD reading during this interval must to be maintained and as well as buffer pH also.
I think the important aspect is not comparing the OD between control (Pyrogallol) and samples. It is obvious that the OD of samples will be greater than that of the control. The important thing is looking at the mean difference per minute. If still the mean difference of samples are still higher, then try to look at your buffer (Tris 0.05M EDTA 0.001M pH 8.5). You can try 20mM pyrogallol.
Best regards
i have done this assay using the Tris Cacodylate buffer ph8.5 and 50 mM pyrogallol.
i need some information regarding the final calculation for the SOD activity.
protein values do tbe used for the unit activity.
- Badges
- Science method