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#1
11-25-2010, 09:25 AM
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 high MW bands are white low MW bands are black?!? I'm doing westerns with an anti ubiquitin Ab on total cell extract. Attached is a picture of my western blot. As I expect ubi to conjugate to proteins of all sizes I expect to get bands of all sizes. And I do, in a way. However, all big proteins come out as negative bands and I do not know why. Anybody ever seen this or have any idea on whats going on? I load 2micrograms of total protein on a 4-20% gradient gel and dilute both 1st and 2nd Ab 1:3500. I have low signal; the blot shown is exposed to a film for 15 minutes. all input would be highly appreciated.  |
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| bands , black , high , low , white |
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