Puzzled
Dr Engelbert Buxbaum
via methods%40net.bio.net
(by engelbert_buxbaum from hotmail.com)
Mon Oct 22 08:25:48 EST 2007
Am 11.10.2007, 07:51 Uhr, schrieb Yvonne Couch
<yvonne.couch from dpag.ox.ac.uk>:
> if I am stimulating
> my cells with an unknown substance and I want to know what cell surface
> receptor it is binding to how do I go about this without systematically
> knocking down every known receptor or running a microarray. Is it
> possible?
You will need a labelled version of your substance, usually radioactive
isotopes are used. With those you can find those fractions of solubilised
cell membranes which have significant binding. A more direct approach is
the use of photoaffinity labels (e.g. azido-groups) where you incubate the
cells, flash them with UV-light and then isolate labelled bands from
SDS-PAGE gels. Note that some natural substances (e.g. ATP) are
photoaffinity labels in their own right. See for example
@book{Scr-84,
Editor= {E.F. Scriven},
TITLE= {Azides and nitrenes},
YEAR= {1987},
PUBLISHER= {Academic Press},
LANGUAGE= {engl}
}
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