Cell-surface molecule control for FACS

Ian A. York iayork at panix.com
Thu Oct 21 09:58:02 EST 2004

In article <ant211151d07zokP at mrc7acorn1.path.cam.ac.uk>,
Mike Clark  <mrc7 at cam.ac.uk> wrote:
>In article <cl6fv4$j03$1 at reader1.panix.com>, Ian A. York wrote:
>> I'm looking for a molecule as a control for flow cytometry.  What I need 
>> is a protein with the following characteristics:
>>  o  Expressed on many (human) tissue types and cell lines
>>  o  Expressed at relatively high levels, so easily detected
>>  o  Expressed at relatively constant levels (i.e. minimally sensitive to 
>> cell cycle stage or stress)
>>  o  Antibodies easily available
>>  o  NOT MHC class I, since that's what I'm trying to control for.

>Ian, you don't say what you mean by many human tissues? 

Mainly adherent cell lines -- Hela, several fibroblast-type lines.  
However, we also will look at lymphoblastoid cell lines, and it would be 
useful (though not essential) to look at the same marker on them.  

Are CD45, CD59, and DAF expressed well on adherent lines?  My vague 
impression was that they were mainly highly expressed on lymphocytes and 
other suspensions type cells.

I'll look into CD18, though.


>Clearly there are pan-leukocyte markers such as CD45 (Leukocyte Common
>Antigen) or the common integrin beta subunit CD18, which will be
>expressed on a large number of cell lines and blood cell types.
>But other markers you might like to consider are things such as  the
>complement regulatory markers such as CD59 and decay accelerating factor
>(DAF) which are expressed quite widely on tissues exposed to complement
>plasma factors. Both CD59 and DAF are GPI anchored if that makes a
>difference to your experiments? 
>Mike                             <URL:http://www.path.cam.ac.uk/~mrc7/>
>M.R. Clark, PhD. Division of Immunology
>Cambridge University, Dept. Pathology
>Tennis Court Rd., Cambridge CB2 1QP
>Tel.+44 1223 333705  Fax.+44 1223 333875

    Ian York   (iayork at panix.com)  <http://www.panix.com/~iayork/>
    "-but as he was a York, I am rather inclined to suppose him a
     very respectable Man." -Jane Austen, The History of England

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