Well-to-well variation observed

Incorrect dispensing and/or experimental setup parameter used.  A fluorescence microplate reader with automatic dispensing should be used for assays requiring precise timing, such as ion channel assays. Faster addition speeds can lead to better mixing of compounds and lower signal variance across the plate. Make sure to follow the recommended experimental setup parameters provided by the instrument manufacturer before reading the plate. It is also important to run the signal test before the experiment.  Different instruments have their own intensity range. Adjust the signal test intensity to the level of 10% to 15% of the maximum instrument intensity counts.

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