I am working with production of cellulolytic enzymes from fungus ( such as Aspergillus sp) and I want to perform Endoglucanase, Exoglucanase and β-Glucosidase Zymograms. I didn't have problems performing the endoglucanase, I used CMC 0.05% and congo red as revelator, I can see 5 different bands from 25 to 40 KDa. But with the Exoglucanase and β-Glucosidase I revised the methods: for exoglucanase, I can use fluorescent substrates such as 4-Methylumbelliferyl β-D-cellobioside in a Native PAGE to see the specific activity. For β-glucosidase I can use another fluorescent substrate such as 4-Methylumbelliferyl β-D-glucopyranoside or Esculin (Kwon et.al
, 1994) to see the band.
The problem is that the fluorescent substrates are expensive, and the delivery times are more than two months.