A cell-based stable assay system for monitoring NF-κB activity and a use thereof

摘要

PURPOSE: Provided are stable system for quantitative determination of NF-kappa B expression in human skin cells and its use in screening skin whitening agents, anti-inflammatory agents or anti-oxidants. The analytical system uses fluorescent substrate to make it very sensitive and simple and is used effectively in screening skin whitening agents, anti-inflammatory agents or anti-oxidants in natural products and organic compounds. CONSTITUTION: The determination system comprises: a fused expression vector, pNF-k B-SEAP-NPT from fusing a gene expressing NF-kappa B transcription reporter, selected from SEAP(secretory alkaline phosphatase), luciferase, beta-galactosidase and GFP(green fluorescent protein), with a selectable marker expression gene chosen from NPT(neomycin phosphotransferase), CAT(chloramphenicol acetyltransferase) and beta-galactosidase, lacZ; transfecting it to skin cell line; and quantitatively determining the expression of NF-kappa B from the above transfected skin cell line(KCLRF-BP-00035) by fluorometry.