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Cell-surface molecule control for FACS

Cell-surface molecule control for FACS - Protocols and Methods Forum

Cell-surface molecule control for FACS - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 10-20-2004, 07:57 PM
Ian A. York
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Default Cell-surface molecule control for FACS



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.

I'm wondering about some of the integrins, perhaps, but I'm not sure which
are widely-enough expressed; but any protein that meets the
characteristics above would be OK.

Ian
--
Ian York ([Only registered users see links. ]) <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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  #2  
Old 10-21-2004, 11:30 AM
Mike Clark
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Default Cell-surface molecule control for FACS

In article <cl6fv4$j03$[Only registered users see links. ]>, Ian A. York
<URL:mailto:[Only registered users see links. ]> wrote:

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

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

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  #3  
Old 10-21-2004, 02:58 PM
Ian A. York
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Default Cell-surface molecule control for FACS

In article <[Only registered users see links. ].cam.ac.uk>,
Mike Clark <[Only registered users see links. ].uk> wrote:


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.

Ian



--
Ian York ([Only registered users see links. ]) <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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  #4  
Old 10-21-2004, 09:10 PM
Kyle Legate
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Default Cell-surface molecule control for FACS

Ian A. York wrote:
Beta 1 integrin has the widest expression pattern. Cell lines that don't
express beta 1 will express beta 3.


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  #5  
Old 11-10-2004, 02:46 PM
Dr Engelbert Buxbaum
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Default Cell-surface molecule control for FACS

Ian A. York wrote:


Housekeeping proteins like Na/K-ATPase, Na/glucose-symporter or the
like?
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