Figure 4. Antibody optimization and validation of negative selection.

A. Each antibody was titrated from 1 to 0.003 µg to determine the amount required for optimal signal to noise ratio for flow cytometry. B. Freshly digested fat pads were subjected to negative selection and the effectiveness of that selection analyzed with flow cytometry by comparing input, or non-selected cells, to selected cells. Single, live cells were gated on CD31 and CD45. After selection, 99.5% of cells were CD31-CD45- compared to 78.4% in the input.