lines through samples - (Sep/23/2004 )
I am having the consistent problem of lines through my samples...I don't know why..there are clear lines down the lanes when I develop and the bands are also "wrinkled" or wavy.
I don't think the gel is heating when I'm running it, nor is it leaking and I'm not overloading.
also, regarding the MAP kinase westerns I can't get it to work even using the positive control..anyone with any advice?
Thanks a lot.
please be a little more specific -
are the bands you see in westerblots "wrinkled", though the ponceau s stain shows the lane itself to be ok? what are the lines you mentioned? protein lines on your gel or blot? at what size? or are the lines on the film? etc.
and the MAP kinase westerns usually are anti phospho-something westerns, yes? i found that anti-phospho antibodies vary greatly in quality and sensibilty, so maybe you would want to change for another antibody from another supplier?