David Hedley writes: >This is following up on Howard's suggestion about DiIC1(5) as a >mitochondrial membrane potential probe to detect apoptosis. In our hands >this gives a large decrease in fluorescence following loss of mitochondrial >membrane potential, either during apoptosis or in response to uncoupling >agents. Because it is red HeNe excited, it should be spectrally compatible >with GFP. > >Our staining protocol for DiIC1(5) is 40nM for 30 min at 37°, excite red >HeNe, collect 675nm. Separation of low MMP cells is greater if you resuspend >in dye-free medium. However, we rarely bother to do this, as we usually see >daylight between high and low MMP populations without resuspending. We have >published several papers using this stain. Most recent (and technically >detailed) is Sheng-Tanner et al., Radiation Research 1998;150:636-647. > >Molecular Probes were thinking of discontinuing DiIC1(5) last time we >ordered some (late 1998), so you might be out of luck. > Sorry, David, I had forgotten you mentioning your use of DiIC1(5) for mitochondrial membrane potential. I'm getting old. Thanks for the references. DiIC1(5) is available from the Acros Organics division of Fisher Scientific (which at one time was the Eastman Organics division of Kodak, from which I bought my original supply): 1,1',3,3,3',3'-Hexamethylindodicarbocyanine iodide, laser grade, 99% (UV-VIS) Quantity:500mg Cat. No:AC41149-5000 Price: I don't know, but probably no more than Molecular Probes charges or charged. -Howard
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