Hi all, I have done Western Blot many times and this is the first time I have seen something like that on my blot!
I ran whole cell lysates on 10% bis-tris gel and subsequently did a Western Transfer at 500mA for 1.5h. When I did Ponceau S after the transfer, the proteins on the blot had discrete bandings. I then probed with my primary antibody at 4oC overnight, and this is an antibody I have used numerous times so I doubt it is the antibody. The only thing I can think of might be that the Western transfer buffer has been used multiple times.
I have attached an imaged. Would love to hear what you guys think!
sad blot.bmp 4.5MB 315 downloads