TNF alpha and IL 1b Western Blot - (Sep/15/2012 )
Hello im trying to do western blot for TNF alpha and for interleukin 1 beta in rat gastric tissue, both proteins are 17Kda proteins,
In the most of the cases I obtain non- specific union or I obtain two strong bands one at the top (250-100 kDa) and another of my molecular marker (aproximadly at 50-25 kDa). I would like some suggestions to remove my nonspecific union (i try milk at diferents conditions and albumin) or maybe it´s my sample. Thansk a lot
What I understood is that all you see are non specific bands and that you are not picking up your proteins of interest. You might want to see whether your proteins of interest have isoforms or whether they can form dimers. The other thing might be the primary antibody you are using. Try using one from a different company and see how the blots look.