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[jwickenden]

I have performed a de-glycosylation reaction using PNGaseF from New England Biolabs as stated on the protocol.
One of my proteins has a known N-glycosylation site and shifted down the gel as expected.
However, the other protein which normally migrates as a diffuse band (at around 39kDa) disappeared and a new band was seen at a much smaller size (about 25kDa). The published size of this protein is 39kDa (by aa sequence) so I don't understand how de-glycosylation would make it so much smaller? Also it isn't thought to have an N-glycosylation site... Does PNGase F remove non-enzymatic glycosylation chains?

Any help from anyone who knows about glycosylation would be great as I am totally new to this area.

Thanks