Im looking to study bile acid transport via apical sodium codependent bile acid transporter (ASBT) in CaCo-2 cells. I have read several papers with brief descriptions of radiolabled bile acids in a transwell assay.
A) I was wondering if any on has done this via fluorescence instead of radioactivity?
B) I have never done radioactivity. Different paper use different labeled bile acids (i.e. 14C, 3H, ect.)....is there a specific reason?
C) Does anyone have a detail protocol for this assay?
For radioactivity, you will have to deal with a number of regulations including training, regular testing and proper disposal. The radioisotope C14 is a beta emitter and beta emissions are easily quenched by the air and even lab gloves. Tritium on the other hand is gamma emitter and gamma particles are higher energy and not as easy to quench. In terms of experimental development, you won't need to deal with the regulatory hassles and safety concerns when you do fluorescence. The sensitivity of of fluorescence may not be as good which is one disadvantage.
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