Shreffler:Eosinophil Isolation

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==Overview==
==Overview==
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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]).
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Isolation of eosinophils for migration studies in collaboration with Bryan Hurley
==Materials==
==Materials==
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*15 mL polypropylene tubes
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*
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*Medium
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*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.
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==Procedure==
==Procedure==
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==References==
==References==
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*[[Special:Emailuser/Wayne G. Shreffler|Email Wayne Shreffler through OpenWetWare]]
*[[Special:Emailuser/Wayne G. Shreffler|Email Wayne Shreffler through OpenWetWare]]
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[[Category:ShreffLab]]
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[[Catagory:Shreffler:Resources]]
 
[[Category:Protocol]]
[[Category:Protocol]]
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[[Category:Flow cytometry]]
[[Category:Immunology]]
[[Category:Immunology]]

Current revision

Contents

Overview

Isolation of eosinophils for migration studies in collaboration with Bryan Hurley

Materials

Procedure

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

Discussion

discuss this protocol

References

Contact

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