I seperated lipoproteins(HDL, LDL, VLDL) by ultracentrifugation but I can not progress. I have problem about g(RCF) and hour for ultracentrifuge to seperate HDL subclasses. I followed a article which is I added the link but it is not clear so I need more clear method.
http://atvb.ahajournals.org/content/27/8/1843.long
The publication you linked refers to an older (1981) paper from JLR (Ref #9):
A density gradient ultracentrifugal procedure for the isolation of the major lipoprotein classes from human serum
Did you follow that method and were unable to separate subclasses as reported?
Could you elaborate on problem with RCF and time?
Most of these methods are not very forgiving and require adherence to details - if you are not using the exact same conditions (or nearly the same), you will not be able to achieve resolution.
Dear Dr Merve,
I have separated the 3 classes of lipoproteins, please find the details about time, procedure, separation, and also oxidation of the 3 classes in the publication attached. If you still have a problem with RCF time please back to me. The methods are straight forward but need some training and practice. I hope this may help
All the best
Thesis Chapter 2-TO INVESTIGATE THE ANTIOXIDANT PROPERTIES OF DIETA...
Dear Prof. Abdel-Moemin,
Firstly, Thank you very much for your interest. I have also seperated the 3 classes of lipoproteins by a different method. But I could not progress. My target is seperating HDL subfractions (like HDL 2a, 2b, 3a, 3b, 3c). I have problem about this subject .
All the best
Dear DR. Alam,
The publication I linked refers to an older (1981) paper from JLR (Ref #9): and it is my problem. I have already seperated the classes of lipoproteins by ultracentrifugation but my target is seperating HDL subclasses.
Dear Dr Merve,
The best practice for myself was the separation for the 3 classes. Try to connect Jane Mceneny, Queen's University Belfast, Department Faculty of Medicine, Health and Life Sciences she may help you in this matter.
Good luck
Aly
Dear Prof. Abdel-Moemin,
Thank you very much for your interest. I am going to try to contact with Jane Mceneny.
All the best
You may consider using size-exclusion chromatography. See J Lipid Res. 2012 Sep;53(9):1890-909.
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