DRG neurons' markers
H.Veldman-2 at neuro.azu.nl
Tue Sep 4 07:05:57 EST 2001
celiacruz at netc.pt schreef:
> Hi everyone!
> I am a portuguese Ph.D student and I'm trying
> to stain big and medium-sized DRG neurons with
> NF200 (N52) antibody using the ABC technique.
> The problem is that the antibody is made in
> mouse and I'm using mouse tissue. I have already
> bought the Mouse on Mouse Kit from Vector but
> I am still having problems. I first tryed diluting
> the antibody 1/500 and 1/1000. Since I got
> a lot of background, I tryed diluting it in
> PBST and incubating at 4ºC. Still bad. I've
> tryed blocking with glycine and normal goat
> serum. Worse!
> I'm really running out of ideas. Can anyone
> give me a hint (as soon as possible, please)on
> how to proceed?
I am afraid the only real solution is to use a rabbit polyclonal
antibody against NF200. We have used one that is sold by Sigma (N4142)
with an ABC-method (Vector) on mouse tissue, after blocking with 5%
normal goat serum at a dilution of 1 : 5000 in 2% normal goat serum.
Hope this helps.
Laboratory for Experimental Neurology (NMZ)
University Medical Center Utrecht (AZU)
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