Phenotyping of apolipoprotein E by immunoblotting in immobilized pH gradients
- 1 January 1991
- journal article
- research article
- Published by Wiley in Electrophoresis
- Vol. 12 (1) , 59-63
- https://doi.org/10.1002/elps.1150120111
Abstract
An immunoblotting method for the determination of apolipoprotein E (apoE) phenotypes has been developed. Delipidated plasma proteins are focused in an immobilized pH gradient, and transferred to polyvinylidene difluoride (PVDF) membranes. ApoE isomorphs are identified by immunoperoxidase staining. The method allows reproducible assignment of apoE phenotypes without isolation of triglyceride-rich lipoproteins. Only small amounts of serum are required. There are several important steps in the procedure: (i) delipidation is indispensable, (ii) carrier ampholytes have to be added to the gels and to the sample buffer, and, (iii) on immunostaining, polyvinylidene difluoride membrane provide an excellent signal-to-background ratio.Keywords
This publication has 25 references indexed in Scilit:
- Apolipoprotein E polymorphism in health and diseasePublished by Elsevier ,2004
- Apolipoprotein E genotyping using the polymerase chain reaction and allele-specific oligonucleotide probesBiochemical and Biophysical Research Communications, 1988
- Native protein blotting after isoelectric focusing in fabric reinforced polyacrylamide gels in carrier ampholyte generated or immobilized pH gradientsElectrophoresis, 1988
- Isoelectric focusing of apolipoproteins in immobilized pH gradients: Improved determination of apolipoprotein EphenotypesElectrophoresis, 1988
- Apolipoprotein E2-Christchurch (136 Arg----Ser). New variant of human apolipoprotein E in a patient with type III hyperlipoproteinemia.Journal of Clinical Investigation, 1987
- Apolipoprotein E phenotyping from serum by Western blottingElectrophoresis, 1986
- Avoiding liquid exudation on the surface of rehydrated gels used for hybrid isoelectric focusing in carrier ampholyte supplemented immobilized pH gradientsElectrophoresis, 1986
- Apolipoprotein EBethesda Isolation and partial characterization of a variant of human apolipoprotein e isolated from very low density lipoproteinsBiochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, 1984
- Identification and characterization of mixed disulphide complexes of e apoprotein in high density lipoprotein of subjects with acute alcoholic hepatitisBiochemical and Biophysical Research Communications, 1979
- Polymorphism of apolipoprotein EClinical Genetics, 1978