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Sanquin’s enzyme-linked immunosorbent assay is a sandwich type immunoassay. The kit contains microwell strips coated with highly avid monoclonal antibodies.
Test samples, calibrator and control sera are incubated in the wells. The IgG subclass to be determined binds to the solid phase and non-bound IgG is removed by washing.
Next peroxidase-conjugated anti-human IgG antiserum is added and non-bound conjugate is removed by washing. After incubation with substrate solution (ABTS) and H2O2 the reaction is stopped with an acid buffer.
The green coloured reaction product is measured by absorption and the concentration of the IgG subclass is calculated relatively to the values of a reference curve.
The IgG subclass concentrations in the calibrators were determined using a calibrator derived from the WHO 67/97 reference preparation.
The kit contains 48 tests, including calibrators and controls, for each of the four subclasses:
• 2 precoated microtiter plates
• HRP conjugated a-IgG antibodies
• standard and control sera
• wash, dilution, substrate and stop buffers
• ABTS and hydrogen peroxidase substrate
Product number of the kit is M1551.
510(k) number: 943098
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