Phagocytosis Assay with Biotinylated Protein
Corresponding Organization : Cornell University
Other organizations : The University of Texas Medical Branch at Galveston, Philipps University of Marburg, University of Southampton
Variable analysis
- Dilutions of guinea pig serum (10-fold serial dilutions, starting at 1:1000 in PBS 2% BSA)
- Phagocytic activity, determined by the area under curve of the MFI (beads positive cells x mean MFI FITC)
- Fluorescent NeutrAvidin beads (Thermo Scientific)
- Biotinylated GPC-I53-50A protein (10μg/5μL beads suspension)
- Incubation of beads with serum for 2 h at 37°C
- THP-1 effector cells (monocytes; ATCC, 5x10^4 cells per well)
- Incubation of beads and cells for 5 h at 37°C
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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