Isolation and purification of rat acute-phase alpha 2-macroglobulin.
Acute-phase alpha 2-macroglobulin was highly purified from the serum of rats in which this protein had been induced 48 h previously by the injection of croton oil, an inflammatory agent. The isolation protocol involved two non-denaturing steps; first, separation according to molecular weight by gel filtration on Ultrogel AcA 22 and second, negative affinity chromatography which bound contaminating proteins to the column while allowing acute-phase alpha 2-macroglobulin to pass through. Several criteria were used to assess the purity of acute-phase alpha 2-macroglobulin, after which the protein by mass determination and by two different protein assays. Pure rat acute-phase alpha 2-macroglobulin was used to produce a monospecific antiserum and to calibrate a secondary standard of rat acute-phase serum by developing and characterizing rocket immunoelectrophoresis assay.
D000818:Animals; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003436:Croton Oil; D007122:Immunoelectrophoresis; D007249:Inflammation / Q000097:blood / Q000139:chemically induced; D008297:Male; D051381:Rats; D000511:alpha-Macroglobulins / Q000302:isolation & purification*
D000511:alpha-Macroglobulins; D003436:Croton Oil