Isolation and characterization of mouse C1q

J Immunol Methods. 1980;36(2):159-71. doi: 10.1016/0022-1759(80)90040-x.

Abstract

C1q was isolated from mouse serum and ascites fluid by absorption onto human IgG-coated latex beads followed by separation on 3-10% exponential gradient sodium dodecyl sulfate (SDS) polyacrylamide gels. Mouse C1q was also purified by low ionic strength precipitation of mouse serum. The purified C1q was heat-labile (56 degrees C, 30 min) both structurally and functionally, contained 4.3% hydroxyproline, 1.38% hydroxylysine, and 18.5% glycine, had an apparent molecular weight of 380,000 daltons, and reconstituted the hemolytic complement activity of C1q-depleted mouse serum. The negatively stained ultrastructural appearance of this purified material consisted of 6 globular units connected by strands. These data demonstrate that mouse C1q structurally and functionally is similar to human and rabbit C1q. A portion of polyacrylamide gel containing mouse C1q was injected into rabbits resulting in the production of monospecific antisera against mouse C1q. Thus, this procedure is a new, rapid and simple method to obtain monospecific antisera against mouse C1q.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adsorption
  • Amino Acids / immunology
  • Animals
  • Antibody Formation
  • Ascitic Fluid / immunology
  • Complement C1 / immunology
  • Complement C1 / isolation & purification*
  • Complement C1 / physiology
  • Electrophoresis, Polyacrylamide Gel
  • Hemolysis
  • Humans
  • Immune Sera / immunology
  • Methods
  • Mice
  • Precipitin Tests
  • Rabbits

Substances

  • Amino Acids
  • Complement C1
  • Immune Sera