From owner-drosophila@net.bio.net Fri Jan 28 22:00:00 1994 Path: biosci!bloom-beacon.mit.edu!usc!howland.reston.ans.net!vixen.cso.uiuc.edu!moe.ksu.ksu.edu!kuhub.cc.ukans.edu!husc-news.harvard.edu!hubio2.harvard.edu!hdeng Newsgroups: bionet.drosophila Subject: myc-tag staining Message-ID: <27JAN199420330886@hubio2.harvard.edu> From: hdeng@hubio2.harvard.edu (HAIYAN DENG) Date: 27 Jan 1994 20:33 EST Distribution: world Organization: Harvard University Nntp-Posting-Host: hubio2.harvard.edu News-Software: VAX/VMS VNEWS 1.41 Lines: 15 Hi everyone, I am trying to do localization study for a protein of interest in Drosophila embryo. The experiment is to generate a stable germ-line transformant to express in vivo the protein with myc-tag at the N-terminus. After the embryo is fixed, I stain the embryo with mAb 9E10 and subsequently with conjugated anti-mouse antibodies, followed by fluorescence microscopy. But I've been having difficulties with this experiment in that the fluorescence signal is extremely, although I can see fairly strong signal with western blot. Any suggestion on how to improve the fluorescence localization experiment will be greatly appreciated. Many thanks. Haiyan .