Use of Non-Denaturing Gel for Activity Staining/Zymogram.

Protein retains their intact native structure when electrophoresis is performed in non-native condition. Thus, it is possible to see activity of protein band in the gel. For example, from a mixture of protein if we have to identify a novel protein showing lactate dehydrogenase activity, what will be experimental process?

After native gel electrophoresis, gel is removed from the gel and soaked in enzyme activity buffer. Lactate and nitroblue terazolium compound is spread on the gel and kept for some time. Wherever LDH enzyme is located, it will produce a coloured farmazan product. Since farmazan compound is insoluble it will not diffuse and make a sharp band.
Similar, activity staining methods are developed for several enzymes.