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. 2007 Dec 10;76(3):1314–1318. doi: 10.1128/IAI.01025-07

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FIG. 3. — Anti-hepta-serotype HCR blocks the binding of HCR/A and HCR/B to GT1b. Gangliosides (0.1 μg in 100 μl methanol) were added to individual wells of an ELISA and incubated overnight at RT. The plates were washed with PBS and blocked for 1 h at RT with 2% (wt/vol) BSA in binding buffer. The plates were then washed and incubated for 1 h at 37°C with the indicated HCR alone (▪), with preimmune sera (□), or with mouse anti-hepta-serotype HCR sera () in 1% (wt/vol) BSA in Tris-buffered saline. Following a washing step, the plates were incubated with an anti-3 FLAG M2 monoclonal IgG-HRP conjugate (1:12,000) in binding buffer. The plates were washed and then incubated with TMB as the substrate. Reactions were terminated with 0.2 M H₂SO₄. Absorbance at 450 nm (A450) was read on a Victor 3V plate reader.

Inline graphic — Anti-hepta-serotype HCR blocks the binding of HCR/A and HCR/B to GT1b. Gangliosides (0.1 μg in 100 μl methanol) were added to individual wells of an ELISA and incubated overnight at RT. The plates were washed with PBS and blocked for 1 h at RT with 2% (wt/vol) BSA in binding buffer. The plates were then washed and incubated for 1 h at 37°C with the indicated HCR alone (▪), with preimmune sera (□), or with mouse anti-hepta-serotype HCR sera () in 1% (wt/vol) BSA in Tris-buffered saline. Following a washing step, the plates were incubated with an anti-3 FLAG M2 monoclonal IgG-HRP conjugate (1:12,000) in binding buffer. The plates were washed and then incubated with TMB as the substrate. Reactions were terminated with 0.2 M H₂SO₄. Absorbance at 450 nm (A450) was read on a Victor 3V plate reader.