Posted in the Western Blot forum but I haven't received any replies, so I thought I would post my question here as well.
I have been unable to get real bands for my ladder- just ghost bands. I use 10 ul (the recommended volume) in 4-20% mini gels and transfer for 1 hr at 100V. I have tried decreasing my secondary concentration, increasing blocking time/concentration, switching to 5% BSA blocking solution, and decreasing ECL substrate incubation time- still no ladder. What else can I try? I'm new to WB so any suggestions would be appreciated.