Mouse C4 (Ss): three-step purification of its C4c fragment and production of a monospecific antiserum.
We report here that a large fragment of mouse C4 (C4c) can be easily purified from serum with good recoveries, and used to produce a monospecific antiserum. The method of isolation of C4c is based on the observation that C4 is easily activated and fragmented in mouse serum. The fragments bind to the C4-binding protein (C4-bp), a newly described macromolecular component of the complement system. The high m.w. complexes were precipitated by dialysis of the serum against 0.1 M acetate buffer, pH 5.0, and purified by passage of the dissolved precipitate through a Sephadex G-200 column. The complexes were dissociated at high salt concentration, and the C4 fragments were isolated by passage of the mixture through a second Sephadex G-200 column. One of the C4 fragments (C4c) was used to immunize rabbits, and a monospecific antiserum was obtained, which showed a reaction of identity between C4 and Slp. Therefore the C4c fragment of C4 and the Slp protein are structurally related.
D000818:Animals; D000918:Antibody Specificity; D003181:Complement C4 / Q000276:immunology / Q000302:isolation & purification*; D005260:Female; D007106:Immune Sera; D007123:Immunoelectrophoresis, Two-Dimensional; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D011817:Rabbits
D003181:Complement C4; D007106:Immune Sera