Purification of mouse haptoglobin by antibody affinity chromatography and development of an ELISA to measure serum haptoglobin levels.
Mouse haptoglobin was isolated from acute-phase serum initially by affinity chromatography on haemoglobin-Sepharose. This proved inefficient, but sufficient material was obtained for use as an immunogen. Rabbit anti-haptoglobin antibodies were used as immunoabsorbents to isolate larger quantities of haptoglobin. Subsequently, specific anti-haptoglobin antibodies were prepared by affinity chromatography on haptoglobin-Sepharose. A direct sandwich ELISA for mouse serum haptoglobin was developed, using affinity purified reagents. The working range of the haptoglobin standard curve was 0.02-0.5 microgram/ml. The reagents did not cross-react with albumin or haemoglobin and the antibody also recognised rat haptoglobin.
Marsden
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CH
Lilly Research Centre Ltd., Windlesham, Surrey, U.K.
Simmonds
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RG
D000818:Animals; D000906:Antibodies; D002846:Chromatography, Affinity / Q000379:methods*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D006242:Haptoglobins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D006454:Hemoglobins; D006801:Humans; D007123:Immunoelectrophoresis, Two-Dimensional; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D051381:Rats; D013045:Species Specificity
D000906:Antibodies; D006242:Haptoglobins; D006454:Hemoglobins