Journal: Frontiers in Immunology
Article Title: Complement Factor H Family Proteins Modulate Monocyte and Neutrophil Granulocyte Functions
doi: 10.3389/fimmu.2021.660852
Figure Lengend Snippet: FHR-1, FHR-5, FHL-1 and mini-FH bind to primary neutrophil granulocytes and monocytes. Binding of 300 nM of each protein (solid lines) to neutrophil granulocytes and monocytes in Ca 2+ and Mg 2+ containing DPBS was measured by flow cytometry. Filled histograms indicate autofluorescence, dotted lines show samples incubated without the proteins added (antibody controls). Binding of the added proteins (solid black line) was analyzed using polyclonal anti-FH Ab for FHR-1, FHL-1 and mini-FH detection, and monoclonal anti-FHR-5 for FHR-5 detection. A representative histogram out of three independent experiments is shown in each case.
Article Snippet: Bound proteins were detected via incubation with the polyclonal goat anti-human FH Ab in DPBS containing fetal bovine serum (FBS, 0.1%, EuroClone, Pero, Italy) for 15 minutes at 4°C, washed and labelled with Alexa-488-conjugated rabbit anti-goat antibody (Thermo Scientific, Waltham, MA) under the same conditions.
Techniques: Binding Assay, Flow Cytometry, Incubation