Fig. (3) Comparison of activities of 2-iodomelatonin determined in the multiplexed assay for the fresh-cells vs. frozen cells and wash vs. nowash conditions. (A) The EC50 values in the FLIPR calcium assay were 17.7, 13.1, 22.0, and 13.8 pM for the frozen cells (NW-FRO) and fresh cells (NW-FRE) in the no-wash assay and the frozen cells (WA-FRO) and fresh cells (WA-FRE) in the cellwash assay, respectively.(B) The EC50 values in the NFAT-beta-lactamase reporter gene assay were 3.60, 3.56, 7.37 and 7.74 pM for the frozen cells (NW-FRO) and fresh cells (NW-FRE) in the no-wash assay and the frozen cells (WA-FRO) and fresh cells (WA-FRE) in the cell washassay, respectively.