Table 1.

ELISA procedures for quantification of total IgA, IgA1 and IgA2 subclasses, IgG, and IgM in human nasal wash

StepQuantification of isotype
IgGIgMTotal IgAIgA1IgA2
1Anti-γ chainAnti-μ chainAnti-α chainRaMbF(ab)2RaM F(ab)2
2Sample or standardaSample or standardSample or standardAnti-A1 MAbAnti-A2 MAb
3Anti-γ chainbio1-cAnti-μ chainbioAnti-α chainbioSample or standardSample or standard
4StreptavidinAP1-dStreptavidinAPStreptavidinAPAnti-α chainbioAnti-α chainbio
5StreptavidinAPStreptavidinAP
  • a Serial threefold dilutions starting at 1:75 (1:15 for IgA2 assay) for nasal wash and at 0.33 μg/ml (3.30 μg/ml for IgA2 assay) for immunoglobulin isotype standard. For a description of the standards, see the text.

  • b RaM, rabbit anti-mouse immunoglobulin.

  • c Biotin conjugate.

  • d Alkaline phosphatase conjugate.