Indirect double sandwich ELISA for the specific and quantitative measurement of mouse IgM, IgA and IgG subclasses

J Immunol Methods. 1989 Apr 21;119(1):117-25. doi: 10.1016/0022-1759(89)90388-8.

Abstract

We have developed sensitive enzyme-linked immunosorbent assays (ELISA) which measure mouse serum heavy chain immunoglobulin isotypes in nanograms per milliliter. In each case the specific isotypic Ig is sandwiched between an isotype-specific antibody used for coating and another isotype-specific antibody coupled to biotin for detection (with alkaline phosphatase coupled to avidin). These methods are simple to perform, specific for each isotype, reproducible with an average coefficient of variation of 5% for IgG1, 3% for IgG2a, 7% for IgG2b, 10% for IgG3, 3% for IgA and 7% for IgM, and at least 100 times more sensitive than radial immunodiffusion. The assays have been used to determine the absolute concentrations of mouse serum heavy chain Ig isotypes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibody Specificity*
  • Enzyme-Linked Immunosorbent Assay* / standards
  • Female
  • Immune Sera / analysis
  • Immune Sera / standards
  • Immunoglobulin A / analysis
  • Immunoglobulin A / standards
  • Immunoglobulin G / analysis
  • Immunoglobulin G / standards
  • Immunoglobulin Heavy Chains / analysis
  • Immunoglobulin Isotypes / analysis*
  • Immunoglobulin Isotypes / classification
  • Immunoglobulin Isotypes / standards
  • Immunoglobulin M / analysis
  • Immunoglobulin M / standards
  • Mice
  • Mice, Inbred C57BL
  • Rats
  • Reference Standards
  • Reproducibility of Results

Substances

  • Immune Sera
  • Immunoglobulin A
  • Immunoglobulin G
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Isotypes
  • Immunoglobulin M