[Protein-analysis] Re: how much immunoreactivity to the band intensity?

Dr Engelbert Buxbaum engelbert_buxbaum at hotmail.com
Sat Jun 24 10:41:21 EST 2006


Ekatherina Touma wrote:

> i have native form and cleaved form of my protein. After some treatment
> native form decreases and cleaved band increases on western blot. But
> the intensity of the cleaved band is much greater that intensity of the
> native band. (same amount of protein loaded). i guess this can mean that
> when cleaved my protein is better exposed to the antibody so
> immunoreactivity if greater....do you have any idea how to normalize
> it??? 

Difficult. Could perhaps be done by purifying the fragments and doing
ELISAs. 

Alternatively, use a different antibody. Preferrably one raised against
the denatured, SDS-PAGE purified antigen. Then this effect should not
come up.


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