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how much immunoreactivity to the band intensity?

how much immunoreactivity to the band intensity? - Protein Forum

how much immunoreactivity to the band intensity? - Protein Forum


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Old 06-20-2006, 07:57 AM
Ekatherina Touma
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Default how much immunoreactivity to the band intensity?



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??? how to know how much of the band is the actual increase in protein expression and how much is due to the change in protein structure? thanx a lot!

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Old 06-24-2006, 03:41 PM
Dr Engelbert Buxbaum
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Default how much immunoreactivity to the band intensity?

Ekatherina Touma wrote:


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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