Catalase is tetrameric protein with a heme group in active site. As EDTA is strong metal chelating agent, there is a probability to affect the activity of the enzyme.
In many commercial assay kit, the homogenizing buffer contains EDTA.
So, should I use EDTA in homogenizing buffer?
Thank in advance
You are right, adding EDTA is probably not a good idea. But nothing prevents you from making your own homogenizatioin buffer without EDTA.
there are kits which has minuscule amount of EDTA (1mM) along with other components like potassium phosphate etc., for catalase activity assays, which dont interfere much with its activity. so you can add such very small amounts in your own homogenization buffer or have additives to give the best results. you can add salts of Zn2+, Ca2+ and Mn2+ but avoid Cu2+ and Mg2+ salts since they have been reported to decrease the catalase activity in many cases.
Best
Thank you all very much for your kind attention.
Would it be scientifically correct if I add EDTA (1 mM) for the western blotting sample preparation?
Is it used as a protease inhibitor (specially for the metalloproteases)???
Lorin Steinhäuser
Thank you very much for this valuable discussion and for the valuable article links.
With regards,
Koner
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