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.
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.
Eap
C B
CB
Clinique psychiatrique universitaire de Lausanne Hôpital de Cery, Prilly-Lausanne, Switzerland.
Baumann
P
P
D000469:Alkaline Phosphatase; D001801:Blood Stains; D014644:Genetic Variation; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D007525:Isoelectric Focusing / Q000379:methods*; D009439:Neuraminidase; D009961:Orosomucoid / Q000235:genetics / Q000302:isolation & purification*; D010641:Phenotype; D014508:Urea
D009961:Orosomucoid; D014508:Urea; D000469:Alkaline Phosphatase; D009439:Neuraminidase