nicotinic receptor western blot
Posted 15 September 2010 - 02:07 PM
Posted 15 September 2010 - 10:12 PM
Posted 05 January 2011 - 02:20 PM
I am using SCG lysates from rat pups to identify a3 nAChR by western blot. The primary antibody I am using is from Santa Cruz. I am resolving the proteins in 10% polyacrylamide gel and transferring the proteins onto a pVDF membrane. I am probing this membrane with primary antibody (0.5ug/ml). The band I am getting is at 100kDa instead of 55kDa. I know that sometimes the a3 nAChR protein can be identified at 60-62kDa region. I do not know what could be the reason. Suggestions regarding why this could be happening will be useful.
In my experience antibodies for detecting nAChR subunits are notoriously crap. They are so crappy, in fact, that an article was actually written about it by Changeux and friends:
J Neurochem. 2007 Jul;102(2):479-92.
Evaluating the suitability of nicotinic acetylcholine receptor antibodies for standard immunodetection procedures.
My suggestion? Find another way to examine your question...