A simple and rapid method for purification of rat haptoglobin for production of antiserum.
Plasma from rats with acute inflammatory response was fractionated on Blue Sepharose CL 6B, to separate haptoglobin from albumin and lipoproteins. Affinity chromatography on Blue Sepharose proved to be a convenient method for crude fractionation of plasma. Pure haptoglobin was obtained by the subsequent affinity chromatography on a rabbit-haemoglobin Sepharose column. Minor amounts of rabbit haemoglobin co-eluted from the haemoglobin Sepharose column but did not influence the monospecificity of the antiserum raised in rabbits. By use of the antiserum in single radial immunodiffusion, the concentration of haptoglobin in plasma from normal rats was measured to be 0.5 g/l.
Wassdal
I
I
Department of Biochemistry, University of Oslo, Blindern, Norway.
Myrset
A H
AH
Helgeland
L
L
D000210:Acute-Phase Reaction; D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D006242:Haptoglobins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D007106:Immune Sera / Q000276:immunology; D007123:Immunoelectrophoresis, Two-Dimensional; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains
D006242:Haptoglobins; D007106:Immune Sera