I need a clear protocol in order to isolate HDL only from blood by using optiprep (iodixanol). Is it possible to isolate HDL without other lipoproteins(LDL,VLDL) and without staining? Because I need it for further investigation. The rotor that will be used is VTi65.1.
the method I used in LDL oxidation paper allows HDL to be lsolated if you use a fractionater to collect fractions from the bottom of the tube up. This method allows a good separation between LDL and HDL.You have the right rotor.....do you have access to beckman fraction colllection system?
Dear Stephanie many thanks for your answer.I do not have access to beckman fraction colllection.
I will collect it by a syringe.as you know the tubes of rotor VTi65.1 are 11.2 ml.
I have to collect 8ml of blood so how many Iodixanol in term of volume and percentage I needed?
if you are buying in your iodixanol at 60% w/v then you need to mix this with plasma to give a 12% solution. In an 11.2 ml tube 5ml of 12% iodixanol/plasma is overlayed with 6% iodixanol/saline....so from 8ml blood you will only get 1 tube.
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