Immunoprecipitation problem

[jabroney8]

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!

[YT2095]

have you tested with another reagent, ninhydrin for instance?

[jabroney8]

No im not sure what nihydrin is or would do. Can Protein A/G fragment when it is heated?