how to separate immunoreactivity and intensity of the band? - (Jun/15/2006 )

ok i have non-degraded and degraded protein over treatment time course....non-degraded decreases and degraded increases....very nice...but intensity of degraded band is much much more than that of non-degraded... ...that could mean that in degraded form my protein reacts much better with the antibody (more exposed) ....so i cant really rely on the intensity of the band that im seeing....any ideas to normalize it? like against a standard on photoshop? what do you think...thanx a lot!

sorry forgot to mention that im talking about western blotting....

Not really sure wht u mean by standards on a photoshop.. but if my guess is rite... you mean employin another protein standard on your blot with a known concentration and testing it.. then compare the intensity with tht of ur samples??? am i rite... i think u could try tht.. good luck..

hi,
what kind of antibodies u r using to detect both degraded and non degraded proteins.
i mean those r monoclonals or polyclonals?
i assume those are monoclonals!!!!

payeli.

ok i have non-degraded and degraded protein over treatment time course....non-degraded decreases and degraded increases....very nice...but intensity of degraded band is much much more than that of non-degraded... ...that could mean that in degraded form my protein reacts much better with the antibody (more exposed) ....so i cant really rely on the intensity of the band that im seeing....any ideas to normalize it? like against a standard on photoshop? what do you think...thanx a lot!

sorry forgot to mention that im talking about western blotting....