The need for IgG2c specific antiserum when isotyping antibodies from C57BL/6 and NOD mice

J Immunol Methods. 1998 Mar 15;212(2):187-92. doi: 10.1016/s0022-1759(98)00015-5.

Abstract

Isotyping and quantitation of murine IgG2a antibodies are widely performed with commercial monoclonal and polyclonal antisera raised against BALB/c IgG2a myeloma proteins. Recently it became evident that inbred mouse strains with the Igh1-b allele do not have the gene for IgG2a and instead express the IgG2c isotype. We show that commercial anti-IgG2a sera cross-react inadequately against IgG2c in immunoblot and ELISA and hence, are not suitable to detect and measure this subclass in mouse strains such as C57BL/6, C57BL/10 and NOD. We have used DNA immunization to generate polyclonal anti-IgG2c serum and demonstrated that it is essential to use IgG2c-specific antiserum to quantify accurately isotypic responses in mouse strains with the Igh1-b allele.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Animals
  • Antibody Specificity*
  • Cross Reactions
  • Enzyme-Linked Immunosorbent Assay
  • Genes, Immunoglobulin
  • Immunoassay / methods*
  • Immunoblotting
  • Immunoglobulin G / blood*
  • Immunoglobulin G / genetics
  • Immunoglobulin Isotypes / blood*
  • Immunoglobulin Isotypes / genetics
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL / immunology
  • Mice, Inbred NOD / immunology
  • Mice, Inbred Strains / immunology*
  • Reproducibility of Results

Substances

  • Immunoglobulin G
  • Immunoglobulin Isotypes