6-OHDA (Oxidopamine) and MPP+ are transported into the neurons via Dopamine transporters, as well as norepinephrine transporters (NET).

At the moment, I'm focused mainly on 6-OHDA, in order to confirm previous studies findings but with my own cell model.

I'm using Desipramine to block NET prior to 6-OHDA treatment. However, I'm unsure as to what concentration I require to ensure that I am efficiently blocking all if not most NETs.

Several previous studies have used concentrations between 0.1 and 1 uM, in various models, but I'm developing my own model.

How can I quantify or provide evidence that NETs are effectively inhibited, and that 6-OHDA is entering the cells only via Dopamine transporters?

I noticed there is a HPLC protocol for Desipramine detection, so I could extract the media, and determine the concentration after 24 hours. However, the limit of detection for Desipramine appears to be 1uM.

Any suggestions on how I can provide evidence that my cells NET are inhibited?

Any help is appreciated,

thank you