In this paper, a sandwich-like composite membrane with super-high thermal stability was designed and fabricated.
For detection, the membrane was incubated with super signal west femto luminol/enhancer solution (thermo scientific) for 3 minutes, and the proteins were visualized using a charge-coupled device camera (BioRad) and quantified with Quantity One (Bio Rad).
After being washed again, the membrane was incubated with Super Signal West Pico Chemiluminescent Substrate (Pierce) for 5 min. The resultant signal was detected using Blue Lite Autorad Film ISC Bio Expresss).
After washing in Tris buffered saline containing 0.05% Tween 20, membrane was incubated with Super Signal West Pico chemiluminescence reagent (Thermo Scientific) and exposed to film (Denville Scientific, Inc., South Palinfield, NJ).
For chemiluminescent detection, the membranes were incubated with Super Signal detection kit (Thermo-Pierce).
At last, membranes were developed with Super Signal West Pico chemiluminsecence reagent (Thermo Scientific) then visualized on X-ray films.
After three washes, NC membranes were treated with super ECL Plus (Applygen) and then the western blot signals were detected by exposure to films.
The membranes were then treated with Super Signal Dura West Extended Duration Substrate (Pierce) and imaged using a cooled charge-coupled camera (UVP).
Following immunolabeling, membranes were washed and reacted with Super Signal chemiluminescence HRP substrate (Pierce Chemical Co).
