Help with Taxol preps

stapletl at mail.nih.gov stapletl at mail.nih.gov
Tue Jun 27 19:42:06 EST 2000


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><
>>
>>
>
>






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