Murine Serum Antibody Titer ELISA

To determine antibody titers in murine serum, HiBond ELISA plates were coated with whole GAS-M1 bacteria, or recombinant M1 protein or isolated M1 HVR [generated as described in (38 (link))] and incubated with serially diluted serum. Captured antibodies were detected using biotinylated goat anti-mouse IgG (Southern Biotech, CAT 1036-08), IgG1 (BioLegend, RMG1-1), IgG2b (BioLegend, RMG2b-1), and IgG2c (Southern Biotech, CAT 1079-08) and streptavidin-HPR (BioLegend, CAT 405210). BSA-coated wells served as negative controls and presented values represent the average of duplicate experimental values with deducted negative control values. All samples were run in duplicate.

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Emami S., Rojas Converso T., Persson J.J, & Johansson-Lindbom B. (2023). Insertion of an immunodominant T helper cell epitope within the Group A Streptococcus M protein promotes an IFN-γ-dependent shift from a non-protective to a protective immune response. Frontiers in Immunology, 14, 1241485.

Publication 2023

biotinylated goat anti-mouse IgG RMG1-1 IgG2b IgG2c

Anti igg Antibodies Antibody Bacteria Elisa Goat Igg1 Igg2b Mouse Murine Recombinant protein Serum Streptavidin

Corresponding Organization : Lund University

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Variable analysis

independent variables

Coating of HiBond ELISA plates with whole GAS-M1 bacteria
Coating of HiBond ELISA plates with recombinant M1 protein
Coating of HiBond ELISA plates with isolated M1 HVR

dependent variables

Antibody titers in murine serum

control variables

BSA-coated wells serving as negative controls

controls

Positive control: Not specified
Negative control: BSA-coated wells

Annotations

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