pertusis toxin -- making a "uniform suspension"

Wolfgang Schechinger wolfsc at ibms.sinica.edu.tw
Wed Aug 8 10:40:25 EST 2001


John, normally adding a little acid or base (you might try 
HOAc or NH4 acetate) might help. (try it on a small aliquot 
first). Calbiochem should be able to give you more advice.

Wo

> I want to use pertusis toxin to inactivate Gi family members in
> transfected COS cells to investigate some signaling pathways.  I
> ordered the pertusis toxin from Calbiochem (cat. no. 516560), and
> it comes as a lyophilized solid to be reconstituted in water. 
> Both the label on the vial AND the catalog description mention
> that I should obtain a "uniform suspension" before using the
> toxin on cells.  I figured that should be easy enough.
> 
> As it turns out (to my surprise), the toxin forms a cloudy,
> particulate suspension when water is added to the lyophilized
> powder.  Usually lyophilized proteins go into solution very
> quickly and easily, but not so here.  I tried vortexing the hell
> out of it, but the clumps of toxin remain.
> 
> I don't know if the toxin would stand up to sonication; and I
> don't want to add a detergent, since that will also get applied
> to the cells when I add the toxin to them.
> 
> 
> Does anyone who routinely works with pertusis toxin have any
> suggestions on how to make a "uniform suspension"?
> 
> 
> John
> 
> hines at pharm.med.upenn.edu
> 
> 


[tiss mezzage wahs broduceRd using TYPO GENERATOR zoffwer]
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Dr. Wolfgang Schechinger
Lab N233 (c/o Dr. Steve Roffler)
Institute of Biomedical Sciences
Academia Sinica
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e mail wolfsc at ibms dot sinica dot edu dot tw
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