Open AccessElectrophoresis of serum lipoproteins (lipoproteinogram) with the Hydragel Lipo + Lp(a)® (Sebia) kit: evaluation of Fat Red 7B staining
Author(s) -
Randa Bittar,
Gautier Pierrat,
Nabil Koujah,
Corentin Poig,
C. Cherfils,
Valérie Fesel-Fouquier,
Laurence Barbot-Trystram,
Dominique BonnefontRousselot
Publication year - 2020
Publication title -
annales de biologie clinique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.167
H-Index - 27
eISSN - 1950-6112
pISSN - 0003-3898
DOI - 10.1684/abc.2019.1510
Subject(s) - repeatability , reproducibility , staining , chylomicron , chromatography , agarose gel electrophoresis , agarose , chemistry , lipoprotein , medicine , pathology , biochemistry , cholesterol , very low density lipoprotein , gene
The lipoproteinogram (or lipidogram) consists in an electrophoretic separation of the main classes of serum lipoproteins. Separation was done in agarose gel using the Sebia Hydragel Lipo + Lp(a)® kit. A repeatability study (n=6) was conducted on 3 sera (1 normolipidemic, 1 hypertriglyceridemic and 1 with a high Lp(a) concentration). The reproducibility was studied on these 3 sera and on an ascites liquid containing chylomicrons, upon 6 days (n=6). A quantitative approach was made by studying areas under the curve and percentages of fractions. In both cases (repeatability and reproducibility), the revelation of the lipoproteins in the gel after electrophoretic migration was made either by staining with Sudan Black (procedure recommended by Sebia), or with Fat Red 7B. Regardless of staining, both repeatability and reproducibility studies show that all lipoprotein fractions were correctly detected at their respective positions, leading to satisfactory interpretations of lipoproteinograms. Our reproducibility study also confirmed a good stability of the fractions over 6 days (storage at +5 ± 3̊C). In addition, the Fat Red 7B staining leads to a shorter technical time (about 40 min) for the gel drying and staining/destaining phases, which allows us to respond more quickly to certain urgent requests such as chylothorax diagnosis.