ab112114 Calcium Assay Kit (Luminometric) - Abcam

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ab112114 Calcium Assay Kit -LuminometricC. Run Calcium Assay:1. Remove the growth medium from the cell plates.Note 1: It is important to remove the growth medium inorder to minimize compound interference with serum orculture media.Note 2: Alternatively, grow the cells in growth mediumwith 0.5 - 1% FBS to avoid medium removal step. In thiscase, 2X coelenterazine- loading solution in 1X assaybuffer is needed.2. Add 100 µL/well (96-well plate) or 25 µL/well (384-wellplate) coelenterazine loading solution (from section B,step 4) into the cell plates3. Incubate the coelenterazine - loading plates at roomtemperature for 3 - 4 hours, protected from light4. Prepare the compound plates with HHBS or the desiredbuffer.5. Monitor the aequorin luminescence intensity by usingthe photon detection system that has an enclosedchamber containing a photomultiplier. The instrumentmust completely exclude outside light.9
ab112114 Calcium Assay Kit -Luminometric7. Data AnalysisFigure 1. ATP Dose Response on CHO-aeq cells. CHO cells stablytransfected with apoaequrin were seeded overnight at 50,000cells/100 µL/well in a white wall/clear bottom 96-well plate. Thegrowth medium was removed and the cells were incubated with100 µL of dye-loading solution using the ab112114 for 3 hours atroom temperature and protected from light. ATP (25 µL/well) wasadded to achieve the final indicated concentrations. The EC 50 of ATPis about 0.8 µM.10
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ab112114 Calcium Assay Kit (Luminometric) - Abcam

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