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Problem with Denative lysis - (Dec/15/2013 )

Dear all,

I am trying purify a protein from inclusion body, so I prepared cell lysates under denative condition by adding urea 8M, but when I run cell lysates on SDS-PAGE, there is no band at all. I'm quite sure that the gel is fine since I checked it several times.

Did anyone has this problem? 

Please give me some advice



how much protein are you loading in each lane?


how are you visualizing the lanes?


do you run standards? if so, how do they look?