Hi! I am working on a project using delipid plasma and was asked to come up with a procedure to prepare it in-house. We are using Heparin (Li) plasma and really was unable to find anything substantial online thus far. Any help is much appreciated!
Laura
I worked on plasma for last 20 years and two basic procedures have worked really well for our lab
- Collect Blood Sample after 12h fasting. Even if 12h fasting sample is not available, never collect it after a meal.
- We use a simple procedure where we centrifuge plasma at high speed (14000g) at -5°C for 30 min. After which you will see a whitish layer of fat around your Eppendorf.
Carefully extract your plasma without fat after this step
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Hello Laura If you are interested in using the Chemical Method for delipidation, then see the steps below
- Mix plasma with chloroform and methanol in a ratio (typically 2:1:1 by volume).
- Vortex the mixture thoroughly to ensure the solvents interact with the plasma lipids.
- Centrifuge the mixture to separate the aqueous (upper) and organic (lower) phases.
- The lipids partition into the organic phase (chloroform), and the aqueous phase contains delipidated plasma proteins.
- Carefully collect the aqueous phase.