In order to investigate the feasibility of screening the NF-κB inhibitors from the Chinese traditional medicine by using dsDNA-coupled microplate (dsDNA-plate),the HeLa cells were treated with small active molecule from the herbs and stimulated with TNF-α for activating NF-κB. The nuclear extracts were then prepared from HeLa cells and the activated NF-κB in nuclear extracts were assayed with dsDNA-plate to find whether small active molecules regulate the activity of NF-κB. The results showed that dsDNA-plate can reveal the NF-κB inhibitory activities of dexamethasone and curcumin which were demonstrated by previous studies. The dsDNA-plate assays also showed that two small active molecules, rescogenine and diosgentin can inhibit the activity of NF-κB. These results suggest that the dsDNA-plate may provide a new platform forscreening NF-κB inhibitors from Chinese traditional medicine.