Shreffler:Eosinophil Isolation: Difference between revisions
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==Overview== | |||
# Under hood, spray down the buffy | |||
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]). | |||
==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== | |||
# 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. | |||
==Discussion== | |||
[[Talk:{{PAGENAME}}|discuss this protocol]] | |||
==References== | |||
==Contact== | |||
*Who has experience with this protocol? | |||
*[[Special:Emailuser/Wayne G. Shreffler|Email Wayne Shreffler through OpenWetWare]] | |||
[[Catagory:Shreffler:Resources]] | |||
[[Category:Protocol]] | |||
[[Category:Immunology]] | |||
Revision as of 10:26, 4 May 2011
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 rhodamine, which can be measured by flow cytometry in the fluorescein channel (spectra).
Materials
- 15 mL polypropylene tubes
- Medium
- DHR-1,2,3 Sigma dissolved in DMSO at 5 mM (500X) and stored at -20°C.
Procedure
- 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.
Discussion
References
Contact
- Who has experience with this protocol?
- Email Wayne Shreffler through OpenWetWare
