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surface staining of THP-1 cell


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#1 maverick3006

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Posted 01 April 2009 - 04:49 PM

Hi
I am trying to perform surface staining for TLRs 2 and 4 on THP-1 cell line. however, i am not getting any staining at all. is the trypsin i am using for cell detachment causing problems in staining?

also, is there a specific protocol for staining THP-1 cells for FACS? please help.

#2 SuMi

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Posted 10 May 2009 - 10:07 AM

Why are your THP-1 cells attached? Did you differentiate them? My THP-1s grow in suspension and so there is no issue with detaching them. I have stained them for TLR2 and TLR4 and they express both proteins quite highly. I used a normal flow staining protocol, probably from BD.

#3 rhombus

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Posted 27 May 2009 - 05:45 AM

Hi
I am trying to perform surface staining for TLRs 2 and 4 on THP-1 cell line. however, i am not getting any staining at all. is the trypsin i am using for cell detachment causing problems in staining?

also, is there a specific protocol for staining THP-1 cells for FACS? please help.



As SuMi states, THP-1 are a suspension culture and only adhere if they are differentiated. I think you have a problem with your cells....have you mycoplasma tested them recently ? .... another cause of phentotypic/morphological changes.

Rhombus




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