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As to the source and structure of the gel I am uncertain. As to how to prevent it .... I think you will find it is the temperature of the centrifugation step. Slight biological variations will answer why they were not all equally affected. I have found that due to the refrigeration units in most modern centrifuges , a programme set to 18oC is INSUFFICIENT. Try 37oc!

George Reid
Beaumont Hospital
Dublin


From: Salter, Simon 
Sent: Friday, May 20, 2011 11:32 AM
To: [log in to unmask] 
Subject: A Friday problem

Dear brain trust,

 

Yesterday I was doing an extraction in blood using 90:10 Hexane:ethyl acetate (4ml). After mixing (pretty vigorously for 10 minutes) and spinning I had a lovely layer of cells at the bottom of my tube and a large layer  of jelly above. The hexane/ethyl acetate mixture had formed a gel with something in the blood. My question is this: what is it in the blood that causes the gel formation and why does it only happen in some samples (it happened in 4 of 40 samples yesterday but normally nothing happens at all, I just end up with a nice layer of hexane/ethyl acetate and a layer of blood at the bottom). All samples are treated the same and mixed at the same time on a vibramixer. I have noticed something similar when extracting with diethyl ether too.

 

My only thoughts is that the hydrophobic solvent crashes the proteins out of solution which then all  bind to each other in a random disorganised manner thus forming a gel. The samples are post mortem and in fluoride oxalate tubes.

 

Anyway, I thought someone out there might have a proper answer.

 

Simon

 

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Please note, archived messages are public and can be viewed via the internet. Views expressed are those of the individual and they are responsible for all message content.
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