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We do detection of various tagged proteins on a fairly routine basis.
We have used the flag mAb from Kodak, which is available in either
purified form or biotinylated. The myc tag is another protein fusion
tag that works well for detection of intracellular proteins. For
methods, see our paper in current issue of Oncogene V. 14, pp1-16,
Wilson et al : Differential subcellular localization, expression and
biological toxicity of BRCA1...
Gary Elliott
AMGEN Flow Cytometry
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Date: 2/10/97 11:52 AM
To: cyto-inbox
From: Matthias Haury
Hello,
I was wondering if somebody has experience detecting fusionproteins with a
"flag" epitope, using a commercial available anti-flag antibody for
intracellular expression...
I haven't tried it, but would like to know if this could possibly work,
especially for somebody who want's to analyse cellcycle expression of a
transfected protein using a flag-fusion construct.
Any details on which mAb to use, minimal expression levels, staining
procedures would be very wellcome.
Thank you very much,
Matthias
_____________________________________________________________________________
Matthias Haury Flowcytometry Dept Immunology Institut Pasteur
mhaury@pasteur.fr Tel: 33 (01) 40 61 31 29 Fax: 33 (01) 45 68 86 39
_____________________________________________________________________________
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Date: Mon, 10 Feb 1997 13:55:55 +0100
To: cyto-inbox
From: Matthias Haury <mhaury@pasteur.fr>
Subject: anti-FLAG mAb intracellular staining...
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web