Antibody binding to nonpermeabilized cells

Beug, Shawn via methods%40net.bio.net (by sbeug from ohri.ca)
Tue Nov 6 14:06:58 EST 2007


Hi all,

 

I am interested in calculating the ratio of intracellular and
extracellular GFP-tagged protein in primary neurons. I am trying to
demonstrate that perturbation of posttranslational modification affects
extracellular/intracellular sublocalization. I was wondering if
immunnostaining for transfected neurons with an anti-GFP antibody (using
a Cy3-conjugated secondary antibody) would be the best way to calculate
this ratio. I have done this before in the past but am unsure if
antibodies would stain intracellular proteins, and as a result, give me
the wrong numbers. 

 

Cheers, Shawn

 


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