alexa fluor

Andy Hoffman andrew.hoffman at
Mon Jan 7 15:08:21 EST 2008

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


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 at
Phone (508) 887 4589
Fax (508) 887 4590

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