Title
A rapid micromethod for apolipoprotein E phenotyping directly in serum
Author
Gaubius instituut TNO
Havekes, L.M.
de Knijff, P.
Beisiegel, U.
Havinga, J.
Smit, M.
Klasen, E.
Publication year
1987
Abstract
A new method for the apolipoprotein E phenotyping has been developed. The method is based on isoelectric focusing of either delipidated or guanidine-HCl-treated serum or plasma in a horizontal slab gel system followed by immunoblotting using either polyclonal or monoclonal anti-apolipoprotein E antibodies as first antibody. Apolipoprotein E phenotyping with this method in 200 serum samples that had been stored at -20 ??C for more than one year gave exactly the same results as obtained with the conventional method based on isoelectric focusing of delipidated very low density lipoproteins isolated from fresh serum followed by protein staining. Compared with the conventional method, the present method is less laborious because ultracentrifugation to isolate VLDL is not needed; it is suitable for large scale screening purposes; it needs only a few microliters of serum or plasma, and can easily be performed with samples with low concentrations of apolipoprotein E. Chemicals/CAS: Apolipoproteins E; Chloroform, 67-66-3; Guanidine, 113-00-8; Guanidines; Methanol, 67-56-1
Subject
Apolipoprotein e
Blood and hemopoietic system
Human cell
Immunoblotting
Isoelectric focusing
Methodology
Micrometer
Serum
Apolipoproteins E
Chloroform
Guanidine
Guanidines
Human
Hyperlipoproteinemia Type III
Isoelectric Focusing
Methanol
Phenotype
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TNO identifier
230373
ISSN
0022-2275
Source
Journal of Lipid Research, 28 (28), 455-463
Document type
article