Isolation and characterization of mouse C1q

Linda M. McManus, Paul K. Nakane

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

C1q was isolated from mouse serum and ascites fluid by absorption onto human IgG-coated latex beads followed by seperation 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°C, 30 min) both structurally and functionally, contained 4.3% hydroxyproline, 1.38% hydroxylsine, 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.

Original languageEnglish (US)
Pages (from-to)159-171
Number of pages13
JournalJournal of Immunological Methods
Volume36
Issue number2
DOIs
StatePublished - Jan 1 1980

Keywords

  • EDTA
  • PAGE
  • PBS
  • RIE
  • SDS
  • Tris
  • VBS
  • ethylenediaminetetracetic acid
  • phosphate-buffered saline
  • polyacrylamide gel electrophoresis
  • rocket immunoelectrophoresis
  • sodium dodecyl sulfate
  • tris (hydrozxymethyl) aminoethane
  • veronal-buffered saline

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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