Antibodies (IgG fraction) are purified from antisera or ascites fluid by differential precipitation with caprylic acid and ammonium sulfate. The polyclonal antibodies are then subjected to immunoabsorption chromatography, and occasionally, affinity purification to remove antibodies that cross-react with other CYP enzymes. Purified antibodies are dialyzed, concentrated if necessary, and diluted to the desired protein concentration in phosphate buffered saline (PBS).

Please refer to our manuscript, "Production and purification of antibodies against rat liver P-450 enzymes," A. Parkinson and B Gemzik, Meth Enzymol, 206:233-245, 1991.

Antibodies are more stable at room temperature than microsomes, however it is best to keep them on ice. After use, re-freeze the remainder at –20˚C. You may also want to dispense into smaller volumes for individual use, to reduce the number of freeze-thaw cycles.

1) Western immunoblotting / ELISA: Enables specific proteins to be detected with antibodies that recognize denatured antigens.

2) Inhibition Studies: Provides information on the function of CYP enzymes. Antibodies help to determine the extent to which a specific CYP enzyme contributes to reactions catalyzed by microsomal preparations.