RE: isolating monocytes - summary

Since the CD14+CD16+ population (as opposed to the CD14++CD16-) is also
CCR2-, you could try first sorting all monocytes via FSC/SSC, then
performing a negative selection for CCR2- cells.  Neither of these steps
necessitates ligation of CD16 or CD14 (or any other target which could lead
to activation), and could give you a pure, non-activated population.

 

Steve

 

-----Original Message-----
From: Ruth Bates [mailto:ruth_louise-bates@hotmail.co.uk] 
Sent: Tuesday, April 29, 2008 11:30 AM
To: cyto-inbox
Subject: isolating monocytes - summary

 

 

Elizabeth R. Simons (esimons@bu.edu
<http://applicationmain_12.4.0080.0327.aspx/?culture=en-GB&hash=2000471368>
) wrote: 


Try magnetic beads loaded with the relevant monoclonal antibody.


Philip Barren (barvp@sbcglobal.net
<http://applicationmain_12.4.0080.0327.aspx/?culture=en-GB&hash=2000471368>
) wrote: 


Check this out

 


www.stemcell.com/product_catalog/rosettesep.aspx


 


their products or tech support should be able to help


you
Nigel Miller (ngam2@mole.bio.cam.ac.uk
<http://applicationmain_12.4.0080.0327.aspx/?culture=en-GB&hash=2000471368>
) wrote:

 

    I frequently sort mono's and gate simply on FSC vs SSC. Works well,cheap
and no messy colours.


Carolina Garcia de Alba Rivas (carogar7@gmail.com
<http://applicationmain_12.4.0080.0327.aspx/?culture=en-GB&hash=2000471368>
) wrote:

 

I've been working with a negative selection kit for CD14+ CD16- monocytes,
this kit it's from Stem Cell laboratory, easy sep technique, I obtain a
90-95% purity and I really haven't observed activation although I haven't
properly analyze the cells for activation. They also have kits for isolation
of other populations and you might find something useful.

 




 


					       
Ruth L Bates BSc(Hons) MSc PhD

"I'd rather die on my feet than live down on my knees"
Wayne Hussey 2007



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