I would like to tag pancreatic beta cell's cilia, as far as I know adenylyl cyclase III is a good marker among a few others. Are there any commercially available antibodies with validated specificity?
Thanks
Hello Gonzalo,
It depends on the kind of cilia you want to stain. For both primary and motile cilia, markers of structural components of the cilium work great : acetylated tubulin, being the most commonly used in the field (clone 6-11B-1, mouse monoclonal). You could also use any other kind of antibodies against tubulin modification (polyglutamylated tubulin, detyrosinated tubulin....). Any antibodies against IFTs would do as well (IFT88 for exemple). For primary cilia (which are the ones you should find in your pancreatic cells), some receptors are highly enriched in the cilium and are thus very good ciliary markers. Arl13b is one of the most widely used marker for primary cilia (There is a good one from the NeuroMab facility at UCDavis). As for Adenylate cyclases, most of them are specific to different cell types. For example, ACIII is widely expressed in neurons' cilia, but I could never see anything in fibroblasts. So unless you know from previous studies that it is expressed in your cells, I wouldn't recommend it.
You can try using anti-DNAI2 antibody which is a polyclonal antibody that recognizes dynein in the axoneme of the cilium. The antibody is commercially available and can be purchased from several companies. The antibody can be used for Western blot analysis and immunohistochemistry.
Hello Gonzalo,
It depends on the kind of cilia you want to stain. For both primary and motile cilia, markers of structural components of the cilium work great : acetylated tubulin, being the most commonly used in the field (clone 6-11B-1, mouse monoclonal). You could also use any other kind of antibodies against tubulin modification (polyglutamylated tubulin, detyrosinated tubulin....). Any antibodies against IFTs would do as well (IFT88 for exemple). For primary cilia (which are the ones you should find in your pancreatic cells), some receptors are highly enriched in the cilium and are thus very good ciliary markers. Arl13b is one of the most widely used marker for primary cilia (There is a good one from the NeuroMab facility at UCDavis). As for Adenylate cyclases, most of them are specific to different cell types. For example, ACIII is widely expressed in neurons' cilia, but I could never see anything in fibroblasts. So unless you know from previous studies that it is expressed in your cells, I wouldn't recommend it.
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