Puzzled

Pow Joshi via methods%40net.bio.net (by pow.joshi from gmail.com)
Fri Oct 12 16:31:03 EST 2007


On 10/11/07, Yvonne Couch <yvonne.couch from dpag.ox.ac.uk> wrote:
>
> Hi all,
>
> I have what sadly may be assumed to be a silly question..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?




The way one can answer you kind of question is have only one variable to
solve: I imagine you should have some proir knowledge of this "unknown
substance" that you are stimulating your cells with?.... if you do, then the
problem gets a little easier: here's what you do: you either tether this
substance which is now known to a matrix, and run your cell extract for the
receptor. 2. Alternatively, do a co-immunoprecipitation, folowed by protein
sequencing; or, (and I am hand-waving here a bit) if you have antibodies
against these substances, probably you could even pick out a clone from the
epression cDNA library.

now, if you the "stimulant remains an unknown substance, I'd imagine, you
would have to go te round about, painful way of first purifying it, at least
partially, before you can get to the answering the receptor Q.

...I understand your dilemma here.... however, here's the silver lining: if
you manage to get some incredibly ingeneous way to get to your problem, you
would end up with a great paper and a patent :))

best,
Pow

Thanks
>
> Yvonne
>
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