The ADCD assay was performed as described (26 (link), 132 (link)). Luminex carboxylated microspheres were coupled with antigens as described for antigen subclass and isotype above. Antigen-coated microspheres (2500/well) were incubated with serially diluted heat inactivated samples (1:10, 1:50, 1:250) at 37°C for 2hours. Guinea pig complement (Cedarlane) freshly diluted 1:60 in PBS was added for 20minutes at 37°C. After washing off excess with PBS 15mM EDTA, anti-C3 PE-conjugated goat polyclonal IgG (MP Biomedicals) (1μg/mL) was added. The beads were then washed and C3 deposition quantified on a MAGPIX (Luminex). The background signal (PBS) was subtracted. Experiments were conducted two independent times. Representative data from one dilution was chosen by the highest signal-to-noise ratio.
Anti c3 pe conjugated goat polyclonal igg
Manufactured by MP Biomedicals
Anti-C3 PE-conjugated goat polyclonal IgG is a laboratory reagent used for the detection and analysis of the C3 complement component. It is a goat-derived polyclonal antibody conjugated with the fluorescent dye Phycoerythrin (PE).
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2 protocols using anti c3 pe conjugated goat polyclonal igg
1
Antigen-Specific Complement Deposition Assay
2
Quantify C3 Deposition on Antigen-Coated Microspheres
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Antigen-coated microspheres were incubated with heat-inactivated sera (37°C, 2 hours). Guinea pig complement (Cedarlane) 1:60 was added (37°C, 20 minutes), then anti-C3 PE-conjugated goat polyclonal IgG (1 µg/mL; MP Biomedicals). C3 deposition was quantified on MAGPIX (Luminex) as detailed above [8 (link)].
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