GFP
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From: | Emma Carter <E.Carter@OxfordBioMedica.co.uk> (by way of Marvin Hanna) |
To: | histonet@histosearch.com |
Reply-To: | |
Content-Type: | text/plain; charset="us-ascii" |
Just to clarify a few things about my GFP.
We are using it as a constitutive marker in a virus, which we are using
for gene therapy, it is our internal control.
I cannot use it as a comparitive control against my other virus, which
has a different label on it, because the detection would be different,
ie fluorescence vs Immuno. So, i want to be able to stain for it, usiing
antibodies.
Now, the antibody that Clontech _used_ to do, worked fine on FFPE
Spheroids though not brilliantly on tissue. It didnt work on frozens.
Clontech then stopped doing it, for reasons of their own, and introduced
a peptide antibody. despite several attepmts, and long conversations
with clontech, i could not get it to work, and in collaboration with
them, concluded that it was unsuitable for immunostaining.
So, now i am down to seraching for something else. I have a polyclonal
anti-GFP from Molecular Probes, which i can get to work, at a push on
transfected cells. I, obviously need to get it working on tissue.
I realise that immuno will not give me a nice green signal, but i dont
want a nice green signal, i want a brown DAB signal!!!!
I hope that makes things a bit clearer....
emma carter
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