Animal cell fractions, such as microsomes, S9, and cytosol, are used in preclinical drug development as in vitro test systems in assays predictive of in vivo behaviors and conditions. These assays specifically provide information on hepatic and extrahepatic metabolism of a drug compound as part of its ADME (absorption, distribution, metabolism, and excretion) profile. or for species comparison to inform later definitive in vitro or in vivo ADME and drug-drug interaction (DDI) studies.
Minipig Liver and Skin Subcellular Fractions for Preclinical Drug Development
Many drug-metabolizing enzymes of the liver and other internal organs are also expressed in the skin. While skin possesses a wide variety of metabolic enzymes, specific enzymatic activity is typically less than 10% of that found in the liver. However, skin plays a significant role in the biotransformation of transdermal xenobiotics. We offer mixed gender and male pooled subcellular fractions, including matching S9, cytosol, and microsomes isolated from Gottingen, Sinclair and Yucatan minipig liver and skin tissue. Other custom preparations of minipig subcellular fractions or tissue may be available upon request.
Minipig liver and skin microsomes are supplied in 250 mM sucrose buffer. S9 and cytosol cell fractions are packaged in a suspension buffer containing 50 mM TRIS-HCI, (pH 7.4 at 4ºC), 150 mM KCI, and 2 mM EDTA.
Available Minipig Subcellular Fractions:
Make sure your microsomes and S9 fractions are properly activated
For best results from assays using microsomes and S9 cell fractions, we recommend using NADPH regeneration to support high metabolism levels by ensuring that the NADPH levels are not a limiting factor in your incubations. Our preferred system, RapidStartTM, uses an enzymatic reaction that converts NADP to NADPH, which is then used as a cofactor by CYPs and oxidized back to NADP, and the cycle continues ensuring stable NADPH levels throughout the incubation. RapidStartTM NADPH regeneration is easily activated by simply adding water. Our easy-to-use formula is the most convenient and flexible NADPH regenerating system you’ll find, allowing users to easily tailor the system’s capability by simply varying the amount of water added. RapidStartTM is perfect for long- and short-term incubations and supports the function of recombinant enzymes and subcellular fractions, allowing you to achieve the most accurate and reliable data during your in vitro assays.
Don’t see what you need? Other preparations can be made available!
If you require subcellular test systems not available through our standard product offerings, Custom Subcellular Preparations can be easily ordered to meet the specific needs of your study.
Our designated Custom Products team regularly prepares and isolates cells and/or subcellular fractions from more than 45 different species and has capabilities to meet unique needs that are unavailable through most vendors– non-standard rodent strains, farm animals, insects, precarious tissue isolations such as adrenal gland or jejunum. Our specialists can carry out over 40 characterization assays, providing the end user with a good foundation for their DMPK assays. See our Custom Products page for examples of past custom preparations or get in touch with a specialist to find out how we can meet your needs.