I have a hydrogel solution with acrylamide, bis-acrylamide (both from Bio-rad), Va-044 from WAKO, PBS, and H20. I fixed my samples for 24 hrs in 4% PFA before placing them in the hydrogel solution for three days. Instead of using a desiccator, I have tried both Peanut oil alone as well as Peanut oil after removing the air in a vacuum from the hydrogel. However, both sets of gel have not polymerized after sitting in a 37 degree water bath for more than three hours. I have tried increasing the acrylamide by 1%, and that has not worked either. Any suggestions on where else I may have gone wrong?