Reply to: RE>anti-FLAG mAb intracellular staining... 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 -------------------------------------- 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 _____________________________________________________________________________ ------------------ RFC822 Header Follows ------------------ Received: by amgen.com with ADMIN;10 Feb 1997 11:36:38 -0800 Received: from castle-smtp.amgen.com (firewall-user@castle-smtp [138.133.19.51]) by www-int.amgen.com (8.8.5/8.8.5) with SMTP id KAA07371; Mon, 10 Feb 1997 10:11:19 -0800 (PST) Received: by castle-smtp.amgen.com; id KAA09757; Mon, 10 Feb 1997 10:11:16 -0800 Received: from flowcyt.cyto.purdue.edu(128.210.60.31) by castle-smtp.amgen.com via smap (V3.1) id xma009729; Mon, 10 Feb 97 10:10:58 -0800 Received: by flowcyt.cyto.purdue.edu (940816.SGI.8.6.9/930416.SGI.AUTO) for cyto-sendout id MAA12353; Mon, 10 Feb 1997 12:32:48 -0500 Received: from mendel.sis.pasteur.fr by flowcyt.cyto.purdue.edu via ESMTP (940816.SGI.8.6.9/930416.SGI.AUTO) for <CYTOMETRY@flowcyt.cyto.purdue.edu> id IAA11366; Mon, 10 Feb 1997 08:25:29 -0500 Received: from cleopatre.pasteur.fr (cleopatre.pasteur.fr [157.99.64.10]) by mendel.sis.pasteur.fr (8.8.5/8.8.5) with ESMTP id NAA15394 for <CYTOMETRY@flowcyt.cyto.purdue.edu>; Mon, 10 Feb 1997 13:56:03 +0100 (MET) Received: from [157.99.140.11] (prelude.metchnikoff.pasteur.fr [157.99.140.11]) by cleopatre.pasteur.fr (8.8.5/8.8.5) with ESMTP id NAA18678 for <CYTOMETRY@flowcyt.cyto.purdue.edu>; Mon, 10 Feb 1997 13:55:59 +0100 (MET) X-Sender: mhaury@mailhost.pasteur.fr Message-Id: <l03020700af24ca814524@[157.99.140.11]> Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" 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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