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I don't know about 18% gels but I regularly detect a 17kDa protein using 4-12% gradient gels (shows up in bottom quarter of gel), then transfer to pvdf using semi-dry transfer, 23V for 60-90min. Maybe trying a lower percentage gel would make transfer easier as 18% is fairly high.
#149519 Western blot question- High % Gel/Low MW Protein
Posted
on 03 February 2013 - 07:49 PM
#149407 Western blot question- High % Gel/Low MW Protein
Posted
on 01 February 2013 - 02:39 PM
Actually the time is probably too long and your voltage is way too high (think how far down the gel a protein would migrate if you ran it at 100 V, now compare to the thickness of the gel...) - the protein will be blowing through the membrane. Either reduce the voltage or the time. You can also place a second membrane behind the first to attempt to catch the protein. PVDF and small pore membranes are better for binding small proteins.
