The High and Low Controls are 10 mM stock solutions of Promazine Hydrochloride and Diclofenac, respectively, in DMSO.
Pipetting devices, DMSO, PBS, UV Plates (Cat # P96UV), and an absorbance plate reader capable of absorbance spectra.
The donor plate membrane is a Hydrophobic PVDF membrane with a pore size of 0.45 µm.
Once solubilized, the lipids should be stored at -20C and used within 6 days.
We recommend against using higher concentrations of DMSO in the PAMPA assay. The spectrophotometric background in the Acceptor solution will increase with higher DMSO concentrations, and the higher concentration of DMSO may alter the membrane and affect the result.
If your compound(s) is/are insoluble in PBS with 5% DMSO, clarify the solution by centrifugation at 13,000xg for 10 minutes then use the supernatant for the Donor solution. For the Equilibrium Standard, mix 80 µL of the new Donor Solution with 120 µL of PBS.
Yes, the kit used to utilize Clonidine as a medium permeability control, but we discovered that this control was subject to oxidative degradation, so we have omitted from the current version of the kit. We continue to look for a replacement medium permeability control, but its absence from the kit doesn’t undercut the performance of the kit.
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Storage: -20°C upon receipt
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