Parallel Artificial Membrane Permeability Assay (PAMPA) Kit
- For quantitative determination of gastrointestinal (GI) membrane permeability of test compounds.
- Convenient. Includes all necessary equipment to run a PAMPA plate.
- Simple and low-cost. Procedure is easy to follow and more affordable than cell-based permeability assays.
- High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day.
- Chemical compounds
- Assay takes 20 hr, hands-on time 1 hr
- 96 tests
- 12 months
- MEMBRANE PERMEABILITY is an important characteristic to determine for evaluating compounds as potential drug candidates. Drugs often need to cross cell membranes in order to reach their target of action and this makes a compound’s ability to passively cross these membranes an important characteristic to evaluate. Permeability can be evaluated by cell-based methods; however, these methods are often expensive and time consuming. Parallel Artificial Permeability Assays (PAMPA) offer researchers a quick, inexpensive method of evaluating the gastrointestinal (GI) permeability of test compounds. BioAssay Systems’ PAMPA Kit provides all the necessary components to run a Parallel Artificial Permeability Assay.
What are the Permeability Controls?
The High, Medium, and Low Permeability Controls are 10mM stock solutions of Chloramphenicol, Diclofenac, and Theophylline, respectively, in DMSO.
What other materials are required to run the assay?
Pipetting devices, DMSO, PBS, UV Plates (Cat # P96UV), and an absorbance plate reader capable of absorbance spectra.
What is the membrane of the donor plate?
The donor plate membrane is a Hydrophobic PVDF membrane with a pore size of 0.45 µm.
My test compounds aren’t soluble at 500 µM in PBS. Can I use higher concentrations of DMSO in the PAMPA assay?
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. The Donor solution is used in place of “500 µM Test Compound”. Since some of the compound was centrifuged out, the new concentration is unknown, so it is instead referred to as Donor Solution.
For the Equilibrium Standard, mix 80 µL of the new Donor Solution with 120 µL of PBS.
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