Im interested in MT1 receptor expression in different cell types and tissues. I did some western blots, and I got some nice bands, but unfortunatelly the bands are not in the same molecular weight as it should be (cca 39 kDa). And worst thing is, that in each tissue or cell type I can see the strong bands in really different molecular weights. Therefore Im searching some paper or anything that could help me to find, what I actually can see in my western blots (forms of MT1 receptor). Does anyone has the same problem? I know that MT1 and also MT2 receptors can create homo and heterodimers and also dimers with other GPC receptors, but the bands I can see are in lower molecular weight than 39 kDa (its about 25 kDa in one type of tissue). So I would be very thankful for any help or your experiences.
In my experience, the commercially available antibodies for MT1 stain around 25 and 50 kDa, and all the antibodies I tested against the knockout mouse stained those same bands. What I did to get around the lack of antibody specificity was to use alternative methods (PCR, melatonin treatment of cell cultures) to show that the receptor was present. Have you compared your samples to negative controls in your blots?
Compare your samples to positive and negative controls (the knockout, if you can get it) and update us on the answer.
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