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. 1979 Jul;37(3):517–527.

Purification of the subunit Clq from the first component of equine complement.

T L McDonald, D Burger
PMCID: PMC1457737  PMID: 91570

Abstract

Initial separation and concentration of Clq from fresh, normal equine serum was accomplished by precipitation in 0.02 M acetate buffer, pH 5.5, at 4 degrees for 24 h. The re-dissolved precipitate was clarified by centrifugation at 80,000 g for 1 h and then dialysed against Tris-HCl buffer (0.05 M, pH 8.0) containing 10-3 M EDTA. The clarified dialysate remained biologically active at 5 degrees for at least 4 weeks. Biological activity of equine Clq was determined by assay of its ability to agglutinate sensitized sheep erythrocytes (EA). Following ammonium sulphate fractionation, Sepharose 4B gel filtration yielded three major peaks. Two protein bands were demonstrated on analysis of the second Sepharose peak by disc acrylamide electrophoresis, pH 8.3. Elution of the protein bands showed EA-agglutinating activity only in the band which migrated furthest toward the cathode. Equine Clq isolated by this method yielded an approximate forty-fold purification in specific activity. Some properties of equine Clq were characterized. Equine Clq was heat-labile, as shown by loss of its EA-agglutinating activity after heating 58 degrees for 15 min. Moreover, storage at 4 degrees and freeze-thaw cycles greatly reduced EA agglutination. Preliminary determination of the sedimentation coefficient indicated that it was comparable to that reported for human and rabbit Clq.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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