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Assay Method Information

Assay Name:  NF-keppaB GFP Assay
Description:  NFκB/Jurkat/GFP transcriptional reporter cell line was obtained from SBI System Biosciences. NF-κB/Jurkat/GFP™ Reporter cells (5×105 cells) were plated at a concentration of 1 million cells/ml into each well of a 24-well plate. TNF-α (5 ng/ml) was added. Compounds were added via serial dilutions to corresponding wells. After 24 hours, 100 μl of the cells were transferred to a well of a Costar® UV plate (96 well, No lid, w/ UV Transparent Flat Bottom, Corning, N.Y., Cat#3635) and the intensity of GFP fluorescence was measured (Excitation 485+/−20, Emission 528+/−20) in a Synergy™ HT Multi-Detection Microplate Reader (BioTech, Winooski, Vt.). The intensities of GFP measured were plotted against the amount of TNF-α.
Affinity data for this assay
 

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Last update November 1, 2007
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