1. Pre-staining Procedure for Electrophoretic Study of Serum-Lipoprotein with acetylated Sudan black B
- Author
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Hitoshi Itoh, Fumihiko Mori, Yukio Ishiwara, Isao Miura, Hiroshi Satowa, and Shigeo Kaizawa
- Subjects
Electrophoresis ,chemistry.chemical_compound ,Chromatography ,Filter paper ,Chemistry ,Ionic strength ,Acetylation ,Lipid moiety ,Sudan Black B ,Staining ,Lipoprotein - Abstract
Paper electrophoresis is one of the most convenient methods for the study of serum lipoproteins. In spite of vast amounts of papers published, there is still many difficulties in the staining prncedure of lipid moiety of lipoprotein, e. g. poor staining, long time pecquired for staining, procedure and strong adsorption of dye to the paper itself. The author tested the pre-staining procedure using acetyl sudan black B prepared by the method of Lillie and Burtner. 0.1ml of saturated solution of the dye in ethanol was added to 1.0ml of serum before application of the serum on filter paper. Both alpha and beta lipoproteins migrate in a usual manner and appear much clear as blue zones against a white background.Examination on the duration of staining, amount of sample, duration of electrophoresis the authors found that the best results could be obtained under the following conditions: buffer; veronal pH, 8.6; ionic strength 0.05; potential gradient; 10volt/cm; amount of sample 0.025-0.035ml/1.5cm; duration of electrophoresis 2-3 hours.Comparing with Swahn's original method, pre-staining procedure reported here was foud to be better in reproducibility, time and cost.
- Published
- 1960
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