THE NEED FOR IGG2C SPECIFIC ANTISERUM WHEN ISOTYPING ANTIBODIES FROM C57BL 6 AND NOD MICE/

Isotyping and quantitation of murine IgG2a antibodies are widely perfo rmed with commercial monoclonal and polyclonal antisera raised against BALB/c IgG2a myeloma proteins. Recently it became evident that inbred mouse strains with the Ighl-b allele do not have the gene for IgG2a a nd instead express the IgG2c isotype. We show that commercial anti-IgG 2a sera cross-react inadequately against IgG2c in immunoblot and ELISA and hence, are not suitable to detect and measure this subclass in mo use strains such as C57BL/6, C57BL/10 and NOD. We have used DNA immuni zation to generate polyclonal anti-IgG2c serum and demonstrated that i t is essential to use IgG2c-specific antiserum to quantify accurately isotypic responses in mouse strains with the Ighl-b allele. (C) 1998 E lsevier Science B.V. All rights reserved.