Shreffler:Eosinophil Isolation: Difference between revisions

Latest revision as of 10:29, 4 May 2011

Isolation of eosinophils for migration studies in collaboration with Bryan Hurley

Under hood, spray down the buffy bags and scissors with 70% EtOH and wait until dry.
Cut open the buffy coat bag and pur blood into a 50 mL Falcon tube.

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 ==Overview==
-Dihydrorhodamine 123 (DHR) is a non-fluorescent, membrane permeable molecule that can loaded into cells to measure oxidative burst. Upon oxidation, it is converted to [http://en.wikipedia.org/wiki/Rhodamine rhodamine], which can be measured by flow cytometry in the fluorescein channel ([http://omlc.ogi.edu/spectra/PhotochemCAD/html/rhodamine123.html spectra]).
+Isolation of eosinophils for migration studies in collaboration with Bryan Hurley
 ==Materials==
-*15 mL polypropylene tubes
+*
-*Medium
-*DHR-1,2,3 [http://www.sigmaaldrich.com/catalog/ProductDetail.do?N4=D1054|SIAL&N5=SEARCH_CONCAT_PNO|BRAND_KEY&F=SPEC Sigma] dissolved in DMSO at 5 mM (500X) and stored at -20°C.
 ==Procedure==
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 ==References==
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 *[[Special:Emailuser/Wayne G. Shreffler|Email Wayne Shreffler through OpenWetWare]]
+[[Category:ShreffLab]]
-[[Catagory:Shreffler:Resources]]
 [[Category:Protocol]]
+[[Category:Flow cytometry]]
 [[Category:Immunology]]