PharmPK Discussion - Cell fractionation / cytosol and membrane isolation

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On 22 Sep 2006 at 13:34:38, "Rob ter Heine" (Aprth.at.slz.nl) sent the message


The following message was posted to: PharmPK

Dear all,

Not sure if  this is the right place to ask:

For an intracellular determination of a drug in peripheral blood
mononuclear cells (PBMCs/lymfocytes) I'm looking for a convenient and
fast method to separate the cytosolic fraction from the membrane
fraction. Does anyone have any experience with this? I don't need the
specific organelles (like mitochondria)... I would like to separate
the cytosol fraction from the remaining cell.

Cheers,

Rob

--
Rob ter Heine, MSc, PharmD
Department of Pharmacology, Slotervaart Hospital
Amsterdam, The Netherlands
E: aprth.-at-.slz.nl
T: +31-20-5124737

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On 25 Sep 2006 at 13:37:33, "shilu singh" (shilu1july.aaa.gmail.com) sent the message


You can use ultracentrifuge, speed 80,000rpm for separation of
cytosolic fraction from membrane fraction.
Sheelendra pratap singh
Dr. Reddy's laboratory
DMPK Dept.

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On 25 Sep 2006 at 17:04:41, "Rob ter Heine" (Aprth.at.slz.nl) sent the message


The following message was posted to: PharmPK

Dear Sheelendra Pratap Singh,

Thank you for your answer. Do you have any reference to this method? I
was also wondering: is cell disruption necessary for this method?

Yours sincerely,

Rob ter Heine

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