Abstract
Alpha(α)6-containing nicotinic acetylcholine receptors (nAChRs) have been implicated in nicotine reward and reinforcement. To date, a commercially available, validated α6 nAChR subunit antibody has not been reported. To evaluate a commercially available neuronal α6 nAChR subunit antibody we performed quantitative western blots on protein from the ventral tegmental area of wild type Sprague Dawley rats. As a first approach to determine the specificity of the antibody, we used a control antigen to block the α6 antibody from binding. Next, we tested the antibody in brain tissue of wild type and α6 knockout (KO) C57BL/6J mice. The α6 antibody was present at a higher than expected molecular weight (63 versus 57 kDa) and the control antigen blocked the α6 antibody, suggesting specificity. However, when we genetically validated the antibody, bands were present in both α6 KO mice and C57BL/6J samples. Taken together, our study highlights the necessity to genetically validate antibodies when possible and we report that a commercially available α6 nAChR subunit antibody is non-specific.