Electroblotting of Immobiline DryPlates applied to identification of human plasma apolipoprotein A-I.
A method for efficient electroblotting of Immobiline DryPlates, allowing subsequent immunological identification of separated proteins has been developed. A thin layer of 1% agarose containing sodium dodecyl sulfate is moulded on the 0.5 mm thick polyacrylamide gel surface after completed electrophoresis. After separation of the agarose-polyacrylamide gel sandwich from the plastic film the rigid gel sandwich could be easily transferred to a nitrocellulose membrane and electroblotting could be performed without adherence of the sticky polyacrylamide gel layer to the membrane. Using this technique human plasma high density apolipoprotein A-I isoforms, over a wide concentration range, could be identified in a heterogeneous mixture, conserving the isoform pattern and band sharpness produced in the immobilized pH gradient experiments.
Holmquist
L
L
King Gustaf V Research Institute, Karolinska Institute, Stockholm, Sweden.
D000178:Acrylamides; D016632:Apolipoprotein A-I; D001054:Apolipoproteins A / Q000097:blood* / Q000302:isolation & purification; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D007202:Indicators and Reagents; D007525:Isoelectric Focusing / Q000379:methods; D008075:Lipoproteins, HDL / Q000097:blood*
D000178:Acrylamides; D016632:Apolipoprotein A-I; D001054:Apolipoproteins A; D007202:Indicators and Reagents; D008075:Lipoproteins, HDL; C059413:Immobiline