Hello everyone,

I was running my 10% gel, I ran it on 90V for 30 minutes followed by 120V for around 1:20, then used semi-dry transfer to nitrocellulose membrane.

When I developed my blot, I got horizontal running lines especially in the upper part of the blot

Anyone knows the cause? is it related to the voltage of running?

Or could it be related to samples temperature? After denaturation of my proteins, I usually keep them at RT. Should I put them on ice instead? because usually I do this step immediately before loading.

Thank you!