I only see the ppt in the medium, not in the cells. The cells behave
exactly as I expect them to in response to the nocodazole. After some time,
there is a lot of the ppt. Much more than I would expect from cellular
kinesin precipitation.
<stapletl at mail.nih.gov> wrote in message
news:8jbhku$1cjb at edrn.newsguy.com...
> Have either of you considered checking if you are precipitating excess
kinesin
> or other microtubule binding proteins (like on a Western)? Are you sure
that
> you are not seeing inclusion bodies?
>> In article <L6V55.21997$dF.854616 at news1.rdc1.il.home.com>, "Centriole"
says...
> >
> >I got the same thing happening with Nocodazole. At first I thought my
cells
> >were contaminated. The interesting thing is, it only seems to
precipitate
> >after more than 24 hours incubation. Before that, I can see no ppt
> >microscopically. I wonder if the pH is an issue here because of that
> >observation. In any event, the expected results still occur (i.e..
> >metaphase arrest with condensed chromosomes).
> >
> >
> >"Ian Harper" <Ian.Harper at sci.monash.edu.au> wrote in message
> >news:394F6E0C.C9F8791E at sci.monash.edu.au...> >> Hi,
> >> A student of ours is using Taxol to disrupt the cytoskeleton, and has
> >> started getting unexpected preciptations forming in the medium after
> >> switching from a pharmaceutical grade to the "sigma" powder. A stock
is
> >> made up either in DMSO or Cremophor El (oil based I think) + Ethanol (
> >> as suggested by supplier), but when the stock is diluted 0.25% vol to
> >> the aqueous, we see needle-shaped crystal forming as taxol goes out of
> >> solution.
> >> Ideas, and protocols would be greatly appreciated,
> >> plz email to me: Ian.Harper at sci.monash.edu.au> >>
> >> Thanks
> >>
> >> Ian.
> >>
> >> ><O><1><O><1><O><1><O><1><O><1><O><1><O><1><
> >> Ian Harper
> >> Confocal Imaging Facility
> >> Dept Biological Sciences
> >> Monash University
> >> Wellington Rd, Clayton
> >> VIC, Australia
> >> Tel: +61-3-99055635, Fax: +61-3-99055613
> >> Email: Ian.Harper at sci.monash.edu.au> >> ><O><1><O><1><O><1><O><1><O><1><O><1><O><1><
> >>
> >>
> >
> >
>
