Abstract
Rat monoclonal antibodies (MAbs) reactive with mouse complement subcomponent C1q were raised applying a principle that requires minute amounts of serum and circumvents purification of serum-derived C1q. The principle involves a) using the high affinity of certain cell-bound antibody isotypes for intercalating C1q from serum of various species, b) selecting such antibodies as are syngeneic to the immunized animal species, thus avoiding the production of antibodies against the intercalating antibodies and c) screening for the anti-C1q MAb in microtiter plates coated with C1q-intercalating MAb isotypes that are heterogeneic to the immunized animal species. We could establish 3 MAbs of IgM subclass, whose reactivity to mouse C1q was shown by ELISA techniques. One of them (RmC13C9) crossreacted with human C1q and was shown by SDS-PAGE and subsequent immunoblotting to recognize the C-chain of mouse and human C1q. The other two MAbs are directed against SDS-sensitive epitopes on mouse C1q and were, therefore, further characterized by native agarose gel electrophoresis and immunohistochemical studies.
References
Apr 1, 1976·The Biochemical Journal·K B Reid, R R Porter
Aug 7, 1975·Nature·G Köhler, C Milstein
Apr 1, 1989·Transplantation·E KremmerJ Mysliwietz
Feb 29, 1988·FEBS Letters·L HsiungK B Reid
May 1, 1988·Molecular Immunology·R HoekzemaC E Hack
Jun 1, 1987·Molecular and Cellular Probes·Y FreiB Stockinger
Jan 31, 1985·Biochemical and Biophysical Research Communications·E KilchherrL K Curtiss
Jan 1, 1985·Advances in Immunology·N R Cooper
Aug 15, 1970·Nature·U K Laemmli
Jan 1, 1982·Hybridoma·T A SpringerF Sanchez-Madrid
Nov 16, 1984·Journal of Immunological Methods·F Stemmer, M Loos
Aug 1, 1983·European Journal of Immunology·N C Hughes-JonesJ C Howard
Jun 15, 1981·FEBS Letters·N Hughes-Jones, S Ghosh
Apr 1, 1980·The Biochemical Journal·D W Mason, A F Williams
Jan 1, 1987·Immunology Today·G J ArlaudJ Gagnon
Citations
Jul 31, 1992·Journal of Immunological Methods·R SchuhS Thierfelder
Aug 1, 1990·Hybridoma·E KremmerM Wasiliu
Aug 6, 1999·Cytokine·C LibertP Brouckaert