A simplistic question from a relative novice flower: Do polyclonal secondaries conjugated to Alexa Fluors (e.g. Alexa Fluor 647 goat anti-mouse IgG (H+L) A21236 - InVitrogen/Mol Probes) that are supposed to be 'highly absorbed' against alternative species (e.g. rat) actually cross-react with rat anti-mouse secondaries at certain concentrations? Can they pick up unstained cells in the absence of other primaries/controls? Lastly, is it suggested to wash after the first round of primary + secondary-alexa, then block with normal mouse Ig as previously mentioned in the CD133 string, before adding other primaries? Thanks so much for your time Andy -- Andrew M. Hoffman, D.V.M., D.V.Sc., Diplomate, A.C.V.I.M. (Large Animal Medicine) Associate Professor Cummings School of Veterinary Medicine Tufts University Director, Lung Function Testing Laboratory Department of Clinical Sciences Bldg 21, Suite 110 200 Westboro Road North Grafton, MA 01536 andrew.hoffman@tufts.edu Phone (508) 887 4589 Fax (508) 887 4590Received on Tue Jan 8 12:18:00 2008
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