From owner-diagnost@hgmp.mrc.ac.uk Wed Feb 5 14:33:19 2003 Return-Path: X-Original-To: diagnost-outgoing Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with SMTP id 910477D140 for ; Wed, 5 Feb 2003 14:33:19 +0000 (GMT) X-Original-To: diagnost-list@hgmp.mrc.ac.uk Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 022547D288 for ; Wed, 5 Feb 2003 14:33:19 +0000 (GMT) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6045) id 531CC7D140; Wed, 5 Feb 2003 14:33:18 +0000 (GMT) X-Original-To: diagnost-submit@hgmp.mrc.ac.uk Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 56C187D0C7 for ; Mon, 13 Jan 2003 15:55:56 +0000 (GMT) Received: from mailbox4.ucsd.edu (mailbox4.ucsd.edu [132.239.1.56]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 3DA637D13C for ; Mon, 13 Jan 2003 15:55:50 +0000 (GMT) Received: from proxy.google.com (proxy.google.com [216.239.35.5]) by mailbox4.ucsd.edu (8.12.3/8.12.3) with ESMTP id h0DF32GS020055 for ; Mon, 13 Jan 2003 07:03:02 -0800 (PST) Received: from sjmm36.sj.google.com (sjmm36.prod.google.com [10.6.39.36]) by proxy.google.com (8.12.6/8.12.3) with ESMTP id h0DF2rib009363 for ; Mon, 13 Jan 2003 07:02:53 -0800 Received: (from news@localhost) by sjmm36.sj.google.com (8.12.6/8.12.3) id h0DF2rej019114 for bionet-diagnostics@moderators.isc.org; Mon, 13 Jan 2003 07:02:53 -0800 To: bionet-diagnostics@moderators.isc.org From: tony.oconnor@may.ie (Tony O'Connor) Newsgroups: bionet.cellbiol,bionet.molbio.methds-reagnts,bionet.diagnostics,bionet.immunology Subject: biotinylated protein for amplification Organization: http://groups.google.com/ Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Content-Transfer-Encoding: 8bit X-Trace: posting.google.com 1042470173 19113 127.0.0.1 (13 Jan 2003 15:02:53 GMT) X-Complaints-To: groups-abuse@google.com NNTP-Posting-Date: 13 Jan 2003 15:02:53 GMT Date: Wed, 5 Feb 2003 14:33:18 +0000 (GMT) Sender: owner-diagnost@hgmp.mrc.ac.uk Precedence: bulk I want to biotinylate a protein to use as an "amplification layer" in an ELISA, as follows: [Solid-phase]-(Ab)-(Ag)-(biotinylated Ab)-(streptavidin)- *{biotinylated protein}n*-(HRP-streptavidin)n I'm doing this to replace a biotinylated anti-streptavidin Ab which is not amplifying the signal. I'm going to try BSA - there's always BSA in the lab :-) Has anyone tried other proteins (adequately-sized with several non-adjacent accessible amino groups) in such a system? Thanks, Tony Tony O'Connor Institute of Immunology NUI Maynooth Ireland tony.oconnor@may.ie .