Dissociating cell sheets

[Anthony Davies]

In article <+74k!hc at lynx.unm.edu>, kim at m44.unm.edu writes:
> Hi group:
> Does anyone have a way of making single cell suspensions of cell monolayers
> that does not trash extracellular protein markers?  Since she is going to be
> using a multiparameter flow analysis, this has to be done by flow rather than
> using something like a Western or RIA.

Gently policeman the cells of TC plastic (commercial scrapers, eg Costar, are 
best) and massage them through a sterile cell strainer (eg Falcon 35 2350).  
This works really well for tightly clumped and/or adherent cells which would 
otherwise be impossible to FACS.
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Anthony Davies                Voice: 865-512361
Institute of Virology         Fax:   865-59962
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UK
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