Complement Deposition Assay Protocol
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Corresponding Organization : Massachusetts General Hospital
Other organizations : Children's Clinical University Hospital, University College London
Variable analysis
- Biotinylated antigens coupled to FluoSphere NeutrAvidin beads
- Incubation of immune-complexes with 10 μl 1:10 diluted plasma samples for 2h at 37°C
- Deposited C3 on beads stained with anti-guinea pig C3-FITC antibody and analyzed on an iQue analyzer
- Incubation of immune-complexes with guinea pig complement in GVB++ buffer for 20 min at 37°C
- EDTA containing PBS (15mM) used to stop the complement reaction
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