I made a fusion construct: my protein (272aa)-linker(9aa)-C terminal eGFP (240aa). The problem is that I see two bands on WB: one with expected size of my protein and another one - 28 KDa, which corresponds to eGFP alone. Restriction profile and sequencing have shown that my protein and linker are there. Where does eGFP alone come from? Also, there are two types of green signal in transfected cells- very bright and dim. Does anyone have any suggestions?
Edited by Holsten, 18 October 2009 - 03:23 PM.