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THP-1 stimulation/collection


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#1 #Tobias

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Posted 04 November 2012 - 04:04 PM

Hello,

I'm attempting to establish a protocol with THP-1 cells stimulated with 1uM PMA for 72 hours (I have also being attempting 48hr) then collecting the cells via scraping and staining them for flow cytometry. However, whenever I collect the cells then try trypan blue counting - I discovered most of the cells to be dead. Visually, they look ok under the microscope before scraping, so I'm wondering if this might be the reason they are dying? (Though I have not previously heard of this before...)
Thoughts?

Thanks.

#2 memari

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Posted 04 November 2012 - 05:23 PM

100 Nano Gram/ml of PMA is enough for differentiation of THP-1.

Edited by memari, 04 November 2012 - 05:24 PM.

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Babak Memari

#3 #Tobias

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Posted 04 November 2012 - 07:01 PM

1uM is also enough (60ng/ml), but I have also tried with 100ng/ml - same issue encountered.




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