Dear Netters,
I have problems with immunodetection on western blot. I am working
on chicory protein extract which contains 10% glycerol, 25 mM K phosphate
buffer pH 7.5, 5uM FAD, 0.5 mM MgCl2, 50 uM TPP and 0.25 mM DTT. The
extract have been tested positively for acetolactate synthase (ALS)
activity. Six month ago, the antibody (rabbit) against ALS has been tested
and was specific. And now, I have one big problem: with or without primary
antibody, the secondary antibody (BIORAD, goat anti-rabbit alkaline
phosphatase coupled one) detects the whole proteins as if it was a naphtol
blue black coloration !
What can I do ?
Have you already had such a problem ?
And did you resolve it ?
I would appreciate any kind of advice or idea that could help me.
Could you mail it or them at: edewaele at pop.univ-lille1.fr
Thank you, Delphine
