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I am writing this for a colleague. He is trying to measure oxidative
burst in macrophages isolated from bovine lymphnode. They are collected via
their adherent properties and he has attempted using dihydrodichlorofluorescein
and dihydroethidium (hydroethidine). He has used phorbol ester to stimulate
the cells and was unable to see any increase in fluorescence compared to
unstimulated cells. The cells are viable. He is contemplating using
phagocytic particles or LPS to stimulate, just in case the cells don't
respond to the phorbol. I would like to hear from anyone with experience
using these dyes and any little quirks/tricks they observed that were
necessary to get them to work. Also please let me know what cells you were
using as well.
Thanks to all who respond,
--- ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Kristi R. Harkins, Lab Director <> Office: 515/294-2472 Cell & Hybridoma Facility <> Lab: 515/294-8504 Iowa State University <> FAX: 515/294-0453 1104 Molecular Biology Bldg. <> E-mail:kharkins@iastate.edu Ames, IA 50011 <> xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
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