The bubbles in the wells is an uncommon issue, but it would not affect the results of your experiment. Prior to the assay, we suggest washing the plate to remove the bubbles as suggested in the following protocol: Wash the plate 3
times with the 1X wash buffer. Discard the liquid in the wells into an appropriate waste receptacle. Then, invert the plate on the benchtop onto a paper towel and tap the plate to gently blot any remaining liquid. It is recommended
that the wells should not be completely dry at any time. Add 300 µl of the 1x wash buffer to each assay well. (For cleaner background, incubate for 60 seconds between each wash.) Repeat steps a-b 2 additional times.