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> Related to the discussion on fluorescences beyond saturation, could anyone
> give me a vague idea of the typical concentration range over which directly
> labelled abs exhibit saturation? Or is this extremely variable depending on Ab
> type, cells, etc?
>
> TIA
>
It's essentially a function of antibody affinity.
[ab]+[ag]<>[abag]
For many of the monoclonal antibodies I work with saturation occurs at
about 5 to 10 ug/ml although some staturate at 1ug/ml and others don't
saturate until about 50ug/ml.
Of course we rarely work at equilibrium.
As an approximate guide the avidity of an antibody is the molar
concentration at which you see 50% saturation. More specifically it
should be the molar concentration of unbound antibody, but for most
antibodies free will be in excess over bound so free can be approximated
to input concentration.
Regards,
Mike Clark, mrc7@cam.ac.uk http://www.path.cam.ac.uk/~mrc7/
-- o/ \\ // || ,_ o Dr. M.R. Clark, Division of Immunology <\__,\\ // __o || / /\, Cambridge University, Dept. Pathology "> || _`\<,_ // \\ \> | Tennis Court Rd., Cambridge CB2 1QP ` || (_)/ (_) // \\ \_ Tel. 01223 333705 Fax. 01223 333875
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