Figure 2.

Monitoring loss of immobilized protein target from BMPA hydrogels during chemical stripping. (A) 9 sets of BMPA hydrogels with immobilized protein were fabricated. These hydrogels were split into 3 triplicate groups consisting of the photobleaching control, the buffer control, and the treatment group. Each group contains n = 9 immobilized protein regions. (B) After each round of incubation, the mean integrated intensity of each immobilized protein region was analyzed. 5.49 mm × 5.04 mm ROIs were defined in each immobilized protein region, from which fluorescence intensity values were summed and normalized to the starting fluorescence intensity. The treatment group, which is incubated in stripping buffer, demonstrates substantial (>50%) loss in protein signal in the first 4 rounds of stripping. By contrast, the buffer control experiments demonstrate a steady decrease in fluorescence (~5% per round) until round 16, at which point the rate of signal loss decreases.