Kevin, In theory this does work, and it will work if you have carefully titrated both the biotinylated antibody and streptavidin together. Many commercial ELISA kits actually do this, and they work quite nicely and reproducibly in my hands. As for doing the necessary work myself to get the ratios correct, I think it would involve a lot of work and you would need to repeat it every time you used a new batch of either the antibody or the streptavidin. Howard probably knows someone who has done it successfully and can tell you how much time it would take. Is it really that much more time for one wash step? Randy Fischer TherImmune Research Corporation 9700 Great Seneca Hwy Rockville, MD 20850 (240) 453-6256 RFischer@therimmune.com > ---------- > From: Kevin Waddick > Reply To: Kevin G Waddick > Sent: Tuesday, April 18, 2000 4:56 PM > To: Cytometry Mailing List > Subject: Biotin-streptavidin staining > > > Is it really necessary to wash cells between incubation with a > biotinylated antibody and adding streptavidin that is linked to a > fluorochrome? That is, doesn't it work as well if the two are added > together, thereby saving a step? I suppose that I could test this > myself, however I would consider limited attempts by me to be > anecdotal. > Others out there may have done actual studies of this and might even > know of tricks to make it work -- assuming that it is not as simple as > I > described. That's what I'm hoping, anyway! > > Kevin G. Waddick, Ph.D. > Parker Hughes Institute > 2657 Patton Road > St. Paul, MN 55113 > >
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