From owner-drosophila@net.bio.net Tue Jul 13 23:00:00 1993 Path: biosci!agate!howland.reston.ans.net!sol.ctr.columbia.edu!usenet.ucs.indiana.edu!fly.bio.indiana.edu!matthewk From: matthewk@fly.bio.indiana.edu (Kathy Matthews) Newsgroups: bionet.drosophila Subject: FRT Stock Info Message-ID: Date: 14 Jul 93 22:58:28 GMT Sender: news@usenet.ucs.indiana.edu (USENET News System) Organization: Biology, Indiana University - Bloomington Lines: 52 Nntp-Posting-Host: fly.bio.indiana.edu Tian Xu has provided the following information for users of his FRT stocks. July 14, 1993 We have some feedback about the FLP/FRT - Marker strains and so we would like to bring three things to the attention of the people who are using these strains. Firstly, people in several labs have told us that the anti-Myc antibody (c-myc, ab-1) from Oncogene Science, Inc. does not work in staining experiments involving MYC marker. We have found that indeed this purified antibody is now working very poorly in staining MYC marker- expressing tissues. We do not know why this is so. Although, we did find that this antibody is unstable under some conditions after we tried to purify it from tissue culture supernatant. The good news is that ascites and supernatants produced from the original cell line (Myc 1-9E10.2) give nice staining results for many people. People could obtain this cell line from American Type Culture Collection (12301 Parklawn Drive, Rockville, Maryland 20852) without charge. People should also keep an eye on the FRT stocks that also carry the FLP recombinase gene. Since both the FLP recombinase gene and its target, the FRT sequences, are present in the same flies, it could have a deteriorative effect. For example, these stocks may accumulate modifiers which interfere with the function of this recombination system. In one case, people have found that they are no longer able to induce clones using one of the FLP/FRT - Marker strains which was working beautifully when they first received it. Since the FLP recombinase gene is under the control of the hsp 70 promoter, one way to keep these strains out of trouble is to maintain them at 18C. In any case, the strains that carry both FRT sequences and the FLP recombinase gene were constructed for the connivance of short-term usage. In a long term, these stocks can be easily regenerated by crossing the FLP gene-carrying chromosomes into the stocks that carry the FRT - Marker chromosomes. Please also note a typographical error in the Table 1 of our Developmental paper (Development 117, 1223-1237. 1993). The abbreviations for the two stocks that carry FRT - NM on the right arm of the third chromosome (w; P[ry+; hs-neo; FRT]82B, P[mini- w+; hs-NM]88C and w hsFLP1; P[ry+; hs-neo; FRT]82B, P[mini-w+; hs-NM]88C) should be 82-NM and 82-NM F instead of 80-NM and 80-NM F. -- Tian Xu -------------------------------------------------------------- Kathy Matthews MATTHEWK@FLY.BIO.INDIANA.EDU Department of Biology MATTHEWK@INDIANA.EDU Indiana University MATTHEWK@IUBACS Bloomington, IN 47405-6801 812-855-5782 .