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IgM binding assay

JP Jason Pinger
SC Shanin Chowdhury
FP F. Nina Papavasiliou
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Flow cytometry samples were prepared as described above (Flow cytometry-based VSG coat replacement analyses), except antibody binding was done in two steps: (1) VSG2-specific polyclonal IgM antisera, diluted 1:100 in mouse serum (Mouse Reference Serum, Bethyl Laboratories, RS10-101). (2) 1:500 dilution of Goat Anti-Mouse IgM-FITC (SouthernBiotech 1021-02) in HMI-9. Samples were incubated for 10 min in each antibody solution and washed once with HMI-9 after each binding step. Control samples labeled with monoclonal anti-VSG2-Alexa647 were prepared using a single antibody incubation step. Forward and side scatter, and dead cell exclusion gating were identical to that described above.

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