Luminescence assay for VEGFR-1 dimerization background signal went up?

10 October 2018 0 5K Report

I am working on a luminescence assay (one of the Glo-kits from Promega) to detect VEGFR-1 dimerization due to PlGF-2 binding. The background signal (control without ligand binding) suddenly went up and the window got smaller. What are some of the possible variables that could cause this? The Glo reagent has been switched but the problem persists. Cells were starved for 4 hours before adding the glo reagent. Cells were plated in a white 96-well poly-D-lysine plate with clear bottom. PlGF-2 was made fresh each day.

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