At 13.20 7/7/99 +1000, you wrote: > >Hi All, > I looked around but can't find any references to.... >Whether the level fluorescence of GFP is related to the temperature at >which it is exposed to an appropriate light source? >My query is related to the lack of detectable GFP signal by flow cytometry, >from a GFP + transgenic mouse (the skin on the tail glows for example) bone >marrow cells. If the temperature is important, then in the mammalian system >the GFP may not be suitbale for the researchers needs. > >Thanks in advance, >Geoff >====================================================================== >Geoffrey Osborne | ____ __ o Ahh! >Flow Cytometry (FACS LAB) | __ `\ <,_ >John Curtin School of Medical Research, | __ (*)/ (*) >Australian National University, | ==============| >CANBERRA, AUSTRALIA. | |--| >Email: Geoff.Osborne@anu.edu.au | |--|... >Phone: 61 2 6249 3694 | >FAX: 61 2 6249 2595 | >-----Surfing the Web?: Try http://jcsmr.anu.edu.au/facshome.html >====================================================================== > >Hi, there is an excellent review by Robert Y. Tsien, in Ann Rev Biochem, 1998, 67:509-544, about GFP, where different GFP mutants, some of them temperature resistan,t are described. From my experience, GFP works perfectly well in mammals cells growed at 37C. Some times to increase the fluorescence signals, we grow the cells at 30C, when it is possible. If GFP doesn't emits, sometimes is due to the fusion protein produced that doesn't allowe the proper GFP folding, Alberto Alberto Alvarez-Barrientos PhD Centro de Citometría de Flujo y Microscopía Confocal Universidad Complutense de Madrid Facultad de Farmacia Ciudad Universitaria s/n 28040 Madrid Spain Tfn. 34 91 3941832 Fax 34 91 3941745
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