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Dot blot anomaly - (Dec/22/2013 )

I carried out a dot blot for various concentrations of my cell lysate and found weak positive staining for my protein of interest at a concentration of 15-25ng. However there was absolutely no staining at higher lysate concentrations (the positive control stained even at 100ng). Can someone tell me why? Thanks!

-Shrubal-

was the positive control a lysate or purified protein?

-Missle-

The positive control was also a whole cell lysate

-Shrubal-