Isoelectric focusing of alpha-1 acid glycoprotein (orosomucoid) in immobilized pH-gradients with 8M urea: Detection of its desialylated variants using an alkaline phosphatase-linked secondary antibody system
- 1 January 1988
- journal article
- research article
- Published by Wiley in Electrophoresis
- Vol. 9 (10) , 650-654
- https://doi.org/10.1002/elps.1150091005
Abstract
A method allowing a clear separation of the different variants of desialylated alpha 1-acid glycoprotein (orosomucoid) has been developed using isoelectric focusing in immobilized pH gradients, supplemented with 8 M urea and 2 % v/v 2-mercaptoethanol. Immunoblotting with two antibody-steps afforded high sensitivity and permitted the detection of about 700 pg of alpha 1-acid glycoprotein in a 20 μL plasma sample diluted 1:28 672. A one year old bloodstain, kept at room temperature, could easily be phenotyped.Keywords
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