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-   -   antibody staining on slides (http://www.molecularstation.com/forum/c-elegans-forum/46644-antibody-staining-slides.html)

Tali Melkman 10-10-2003 06:43 PM

antibody staining on slides
 
Hi!
I am trying to stain both adults and embryos on polylysine coated
slides. I have been putting adults on coated slides and pressing down
slightly such that they burst open at the vulva releasing their eggs.
Then I permeabilize the animals by freeze cracking.
My first problem is that the eggs then sometimes stick to the slides
but the adults most times don't. How do I get the animals to stay on
the slide?
The second problem is that I have high nuclear background that
fluoresces orange on the GFP filter, but has nothing to do with the
fluorophore the secondary antibody is conjugated to. This background
then interferes with staining visualized with FITC. What is this
background? how do I get rid of it?
I would appreciate any suggestions/ideas.
Thanks in advance,
Tali

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