The need for IgG2c specific antiserum when isotyping antibodies from C57BL/6 and NOD mice.
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.
The Walter and Eliza Hall Institute of Medical Research, P.O. Royal Melbourne Hospital, Parkville, Victoria, Australia.
R M
RM
Martin
J L
JL
Brady
A M
AM
Lew
D000483:Alleles; D000818:Animals; D000918:Antibody Specificity; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D005803:Genes, Immunoglobulin; D007118:Immunoassay / Q000379:methods*; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000097:blood* / Q000235:genetics; D007132:Immunoglobulin Isotypes / Q000097:blood* / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL / Q000276:immunology; D016688:Mice, Inbred NOD / Q000276:immunology; D008815:Mice, Inbred Strains / Q000276:immunology*; D015203:Reproducibility of Results
D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes