TY - JOUR
T1 - Isolation and characterization of mouse C1q
AU - McManus, Linda M.
AU - Nakane, Paul K.
N1 - Funding Information:
The authors gratefully acknowledge the helpful comments of Dr. R. Neal Pinckard, the amino acid analyses by Dr. Masaru Imada, and the typing of this manuscript by Cynthia Weaver. This work was supported by Grants AI-09109 and CA-15823 from the National Institutes of Health.
PY - 1980
Y1 - 1980
N2 - 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.
AB - 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.
KW - EDTA
KW - PAGE
KW - PBS
KW - RIE
KW - SDS
KW - Tris
KW - VBS
KW - ethylenediaminetetracetic acid
KW - phosphate-buffered saline
KW - polyacrylamide gel electrophoresis
KW - rocket immunoelectrophoresis
KW - sodium dodecyl sulfate
KW - tris (hydrozxymethyl) aminoethane
KW - veronal-buffered saline
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U2 - 10.1016/0022-1759(80)90040-X
DO - 10.1016/0022-1759(80)90040-X
M3 - Article
C2 - 7430648
AN - SCOPUS:0018895387
VL - 36
SP - 159
EP - 171
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
SN - 0022-1759
IS - 2
ER -