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

Electrophoresis. 1988 Oct;9(10):650-4. doi: 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 microL plasma sample diluted 1:28 672. A one year old bloodstrain, kept at room temperature, could easily be phenotyped.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase
  • Blood Stains
  • Genetic Variation
  • Humans
  • Hydrogen-Ion Concentration
  • Immunoblotting
  • Isoelectric Focusing / methods*
  • Neuraminidase
  • Orosomucoid / genetics
  • Orosomucoid / isolation & purification*
  • Phenotype
  • Urea

Substances

  • Orosomucoid
  • Urea
  • Alkaline Phosphatase
  • Neuraminidase