Hello!

I have samples of proteins extracted from aortic tissue using NE-PER™ Nuclear and Cytoplasmic Extraction Reagents, I have attempted immunoblotting a few times already, but the bands I am obtaining are slightly bigger in size than expected.

A deglycosylation step is probably required, I have found a few protocols where the protein I am looking at is isolated first and then chondroitinase and keratanase digestion follows.

Considering my samples are total protein extracts, could I just incubate them with chondroitinase ABC at 37C for 2h, as suggested below and then carry on with WB?

https://linkinghub.elsevier.com/retrieve/pii/S0304416520303159

Does anyone have experience or suggestions on how to optimise WB with glycosaminoglycans?

Thank you in advance!