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Microheterogeneity of apolipoprotein D as revealed by electroblotting following isoelectric focusing in Immobiline DryPlates
Author(s) -
Holmquist Leif
Publication year - 1992
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150130153
Subject(s) - electroblotting , isoelectric focusing , chromatography , chemistry , apolipoprotein b , agarose , gene isoform , sodium dodecyl sulfate , gel electrophoresis , polyacrylamide gel electrophoresis , electrophoresis , biochemistry , microbiology and biotechnology , cholesterol , biology , enzyme , gene
The microheterogeneity of apolipoprotein D was examined by a procedure involving, in sequences: (i) electrophoresis in an immobilized pH 4–7 gradient in an Immobiline DryPlate‐polyacrylamide gel supplemented with Ampholine pH 5–7, (ii) covering of the gel with sodium dodecyl sulfate‐containing agarose, (iii) electroblotting onto a polyvinylidene difluoride membrane and (iv) immunological identification. Seven isoforms were obtained with partially purified apolipoprotein D. Using this technique the apparent p I values at 15°C for the isoforms were 4.57, 4.67, 4.78, 4.83 and 5.95, 6.06 and 6.19 (SD ± 0.05 for all). Direct staining of the Immobiline DryPlate could not reveal the isoforms of partially purified apolipoprotein D.

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