What method can I use to investigate phosphate transport in living human cells?

09 September 2015 1 10K Report

I'm conducting an analysis of a phosphate transporter and I want to measure its transport capacity in human cells.

I cannot use radioisotopes in this experiment.

So, you can't actually measure activity by Pi uptake... What about comparing pERK by Western blotting after you have added either PTH, FGF23 or secreted Frizzled Protein 4, in the presence of Na containing vs Na-free medium and look for increased internalization of NaPi (-1, 11a or 11b) with and without PD098059. Alternatively look for expression of each of the transporters in the cells and use siRNA to knockdown the one that's present? While the PTH exp would not directly measure influx it would tell you the transporter is present and may be regulated.

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