Hi, I'm looking for a possible reason why I got low yield protein from a Pierce GST spin-column (Thermofisher) after an ON incubation with GST-cleavage enzyme and concentrating in an amicon filter? I used 4 x 0.2 mL- spin columns and each column is capable of purifying max of 2mg and I'm expecting a minimum of 10mg/mL of protein from my original lysate but ended up with 0.559 mg/mL. I do not know at which point I lost my protein, please does anyone have idea about this problem?
Thank you.
Isiaka.