[Neuroscience] Pertussis toxin?

Bill via neur-sci%40net.bio.net (by connelly.bill from gmail.com)
Fri Jul 27 05:19:28 EST 2007


I'm starting to feel really guilty for asking so many questions on
this group, but the answers are often so good I can't resist.

I'm investigating a novel G-protein coupled receptor in cortical brain
slices and I want to see whether it works via Gi, Gs or Gq. I remember
simple pharmacology, If pertussis toxin blocks the response of the
agonist, it's Gi mediated. The methods I found are a bit
heterogeneous, anywhere from 2 to 20 hours of incubation with 2-5ug/mL
of pertussis toxin at 22 to 37 degrees.

That was fine, I could empirically figure out how long I needed to
incubate the slices by using a well established ligand (I was thinking
baclofen), -BUT-

My holding chamber holds about 70mL, to fill that with 2ug/mL
pertussis, that would require 140ug of pertussis. 50ug of pertussis
toxin cost $500 US from sigma (only $170 from List Biological Labs);
so that's $1,500 a day (or $500).

Either way, that is big big money in my books. Can anyone either a)
see a mistake in my logic or b) figure a way around this problem?



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