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Schematic of ELISA-IAA assay design. The bridging ELISA-IAA assay is designed for IAA to bind two differently labeled proinsulin molecules in the fluid phase. The antigen (proinsulin)-antibody (IAA) complex generates a measurable signal through enzyme-substrate reactions, which are detected using a standard ELISA plate reader. mAb, monoclonal antibody; SA, <t>streptavidin;</t> <t>TMB,</t> <t>3,3′,5,5′-tetramethylbenzidine</t> (peroxidase substrate).
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Schematic of ELISA-IAA assay design. The bridging ELISA-IAA assay is designed for IAA to bind two differently labeled proinsulin molecules in the fluid phase. The antigen (proinsulin)-antibody (IAA) complex generates a measurable signal through enzyme-substrate reactions, which are detected using a standard ELISA plate reader. mAb, monoclonal antibody; SA, <t>streptavidin;</t> <t>TMB,</t> <t>3,3′,5,5′-tetramethylbenzidine</t> (peroxidase substrate).
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Schematic of ELISA-IAA assay design. The bridging ELISA-IAA assay is designed for IAA to bind two differently labeled proinsulin molecules in the fluid phase. The antigen (proinsulin)-antibody (IAA) complex generates a measurable signal through enzyme-substrate reactions, which are detected using a standard ELISA plate reader. mAb, monoclonal antibody; SA, streptavidin; TMB, 3,3′,5,5′-tetramethylbenzidine (peroxidase substrate).

Journal: Diabetes

Article Title: Identifying Insulin Autoantibodies With Differential Risk in Type 1 Diabetes With a Novel Bridging ELISA

doi: 10.2337/db25-0696

Figure Lengend Snippet: Schematic of ELISA-IAA assay design. The bridging ELISA-IAA assay is designed for IAA to bind two differently labeled proinsulin molecules in the fluid phase. The antigen (proinsulin)-antibody (IAA) complex generates a measurable signal through enzyme-substrate reactions, which are detected using a standard ELISA plate reader. mAb, monoclonal antibody; SA, streptavidin; TMB, 3,3′,5,5′-tetramethylbenzidine (peroxidase substrate).

Article Snippet: HRP-conjugated streptavidin (Thermo Fisher Scientific) is added and incubated at room temperature for 1 h. After additional washes, the peroxidase substrate 3,3′,5,5′-tetramethylbenzidine (Thermo Fisher Scientific) is added to develop color.

Techniques: Enzyme-linked Immunosorbent Assay, Labeling