Simple quantification of complement factors C3 and C3b using separation byisotachophoresis

Authors
Citation
F. Acevedo, Simple quantification of complement factors C3 and C3b using separation byisotachophoresis, ELECTROPHOR, 20(3), 1999, pp. 469-472
Citations number
10
Categorie Soggetti
Chemistry & Analysis
Journal title
ELECTROPHORESIS
ISSN journal
01730835 → ACNP
Volume
20
Issue
3
Year of publication
1999
Pages
469 - 472
Database
ISI
SICI code
0173-0835(199903)20:3<469:SQOCFC>2.0.ZU;2-R
Abstract
The separation of complement factors C3 and C3b by isotachophoresis in 1% a garose gel followed by immunoprecipitation and quantification is presented. Glycine was used as spacer in a nonequilibrium isotachophoresis (Acevedo, F., J. Chromatogr. 1991, 545, 391-396). Tricine, beta-alanine and Tris were the leading Ion, terminating ion ana counter ion, respectively. After elec trophoresis the gel was incubated in rabbit antihuman complement factor C3c . The amounts of C3 and C3b in the sample were measured by optical densitom etry of the Coomassie Brilliant blue-stained immunoprecipitates in the agar ose gel. The correlation coefficient obtained for the logarithm of the inte grated densitometric measurement vs, the logarithm of the amount of applied C3 was higher than 0.98 in calibration experiments. The extent of compleme nt factor C3 activation is calculated as the ratio between the amount of C3 b and the amount of C3b plus C3 and expressed as percent. The progress of c omplement activation from human blood plasma samples induced by Mg2+ and zy mosan are presented as examples.