To Dave and other Zenon users on the list, I would like to hear more about your experiences using the Zenon labeling kit, particularly whether the Zenon-labeled antibody can be used in conjuction with other mouse mAbs. I have heard that there is a problem with the Zenon anti-IgG1 reagent transferring to other antibodies in the staining mix, even if staining is done in separate reactions with washes in between. I would expect to see this only if the other Abs were also mouse IgG1, but avoiding use of this isotype would rule out alot of Abs. It seems to me that the major utility of this reagent in flow cytometery is in generating a wider spectrum of directly labeled Abs for multicolor analyses. Obviously if the label can jump to other Abs, its applications in flow will be more limited than if it stays put. Thanks, Mandy >I agree that I haven't seen anything useful for immunophenotyping >using the 325 nm line, although I do have a couple more that I will >try in the near future. I know that some people have said that they >can make Alexa 350 work, but it didn't work very well in my hands. > However, the 325 nm line IS useful for exciting aminostilbamidine >methanesulfonate. We use it frequently for dead cell >discrimination. It gives resolution between live and dead better >than 7AAD and almost as good as PI. It is available from Molecular >Probes. > >If you're interested in getting 2 colors from the UV/V range >(besides Indo) see if you can swap your 325 nm for a 405 nm. This >would allow you to use Cascade Blue and Yellow. Talk to your sales >rep to see what you can work out. > >One more tip: If you start to use more of these "exotic" fluors, >you'll be hard pressed to find them conjugated to your Abs of >choice. Another reagent from Molecular Probes that has really >opened some doors for us is their Zenon Ab conjugation kit. They >have a wide range of fluors available. They have kits to conjugate >mouse IgG1 and rabbit Ig and I believe they will also have kits for >mouse IgG2a and 2b in the near future. It's good stuff! > > >Happy flowing, > >Dave > > >David McFarland >GlaxoSmithKline > >-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Mandy Cromwell, Ph.D. Department of Immunology New England Regional Primate Research Center Harvard Medical School One Pine Hill Drive Southborough, MA 01772 office:(508)624-8022 FAX:(508)786-1482
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