CH50 ELISA
Immunoenzymatic colorimetric method for quantitative determination of complement functionality.
Test Principle
The complex β-galactosidase/anti-β-galactosidase, is solubilized by serum trough the deposition of C3b molecules. The formation of C3b quantity necessary for the solubilization is mediated by alternative path-way, but it is accelerated from activity of C3-convertase by classic way.
The quantity of complex β-galactosidase dissociated, detectable by enzymatic activity in the supernatant at the end of reaction; represents the capacity of serum to form C3b molecules. The o-nitrophenil-galactopiranoside (o-NPG) is used as substrate and the reagent product (o-nitrophenol) is read at 420nm (or 405 nm).
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References
- Miller G.W, Nussenzweig V.: A new complement function: solubilization of antigen-antibody aggregates. PNAS, 72, 418 – 1975.
- Takahaschi M., Takahaschi S., Brade U., Nussenzweig V.: Requirement for the solubilization of immuno-aggregates by complement. J. Clin. Invest. 62, 349 – 1978.
- Migliorini P, et al. J. of Immunological Methods, 77 119-130 (1985)
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