CAT Enzyme Assay System With Reporter Lysis Buffer

Chloramphenicol acetyltransferase (CAT), encoded by a bacterial drug-resistance gene, inactivates chloramphenicol by acetylating the drug at one or both of its two hydroxyl groups. This gene is not found in eukaryotes, and therefore eukaryotic cells contain no background of CAT activity. This characteristic, along with the ease and sensitivity of the assay for CAT activity, has made the CAT gene one of the first reporter genes used for studies of mammalian gene expression.

Linkage of putative regulatory sequences to the appropriate pCAT ® 3 Reporter Vector and subsequent transfection allows for the efficient assay of CAT activity in cultured cells.

CAT activity may be monitored by two alternative methods in the CAT Enzyme Assay System. The most rapid, sensitive and convenient of these assays is based on liquid scintillation counting (LSC) of CAT reaction products. CAT activity also can be analyzed using thin-layer chromatography (TLC). This method is more time-consuming than the LSC assay but allows visual confirmation of the data.

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