jabroney8 Posted November 25, 2008 Share Posted November 25, 2008 Hello Im trying to do immunoprecipitation on apolipoprotein A1 in human plasma. after transfer, i always ponceau stain the blot to check for transfer efficiency. I usually see a band aroung 28kDa, which is most likely APOA1, another band at 50, which is most likely Protein A/G, and another around 60kDa, which Im not sure what it is. Anyway, when i develop the blot colorimetrically, I see a train of about 8 lanes 1KDa apart from about 34kDa-26kDa that were not visible with ponceau stain. Im confused because the bands are very intense, do NOT show up on a ponceau stain, and are probably masking my protein of interest. i dont know if this train is protein or something else...anyone have any ideas? what can i do to get around this? THANKS! Link to comment Share on other sites More sharing options...
YT2095 Posted November 25, 2008 Share Posted November 25, 2008 have you tested with another reagent, ninhydrin for instance? Link to comment Share on other sites More sharing options...
jabroney8 Posted November 26, 2008 Author Share Posted November 26, 2008 No im not sure what nihydrin is or would do. Can Protein A/G fragment when it is heated? Link to comment Share on other sites More sharing options...
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