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Nuclear extracts prepared from untreated U-937 and treated HeLa (TPA, 10 min) cells are diluted to 0.625 μg/well and assayed using the TransAM Elk-1 Kit. The ratio of the signals from the treated cells over the untreated must be above 4. Lot No. 05504001 was developed for 8 minutes. It gave a ratio of above 4 (Figure 1). The endogenous level of Elk-1 expression, and this ratio may vary depending on the cell type tested and the treatment used. TransAM Elk-1 Kits are also tested for specificity in detecting Elk-1 activity. TransAM Elk-1 assays are performed in the presence of an excess of oligonucleotide containing a wild-type or mutated Elk-1 consensus binding site (Figure 2). At 10X excess, the wild-type oligonucleotide prevents Elk-1 binding to the probe immobilized on the plate. Conversely, the mutated oligonucleotide has no effect on Elk-1 binding.