RRC ID 44048
Author Ukaji T, Kai O.
Title Isotype analysis of gerbil-mouse heterohybridomas by RT-PCR.
Journal J Immunol Methods
Abstract We designed primer sets specific to the immunoglobulin (Ig) heavy-chain constant region (IGHC) genes in Mongolian gerbil (Meriones unguiculatus) to amplify five gerbil IGHC cDNA sequences, Cμ, Cγ1, Cγ2, Cε, and Cα. Five gerbil-mouse heterohybridomas B11D2(C2), B11E2(D5).M, B5-3, D5, and C11 respectively expressed Cγ1, Cμ, Cγ2, Cγ2, and Cγ1. In contrast, a commercial isotyping kit for mouse Igs identified Cγ1, Cμ, Cγ3, Cγ3, and Cγ1, respectively, misidentifying gerbil IgG2 as IgG3 by cross-reactivity with anti-mouse IgG3 polyclonal antibody. These primer sets will allow the accurate estimation of gerbil Ig classes and IgG subclasses. These results from three gerbil strains indicate that the primer sets can be used for isotype analysis of gerbil mAbs and for evaluation of humoral immunity.
Volume 386(1-2)
Pages 108-11
Published 2012-12-14
DOI 10.1016/j.jim.2012.09.001
PII S0022-1759(12)00269-4
PMID 22974835
MeSH Animals DNA / analysis DNA Primers / genetics* Gerbillinae / immunology Hybridomas / immunology* Immunoglobulin Constant Regions / genetics* Immunoglobulin Isotypes / genetics* Mice Real-Time Polymerase Chain Reaction / methods Sensitivity and Specificity Species Specificity
IF 1.901
Times Cited 2
WOS Category BIOCHEMICAL RESEARCH METHODS IMMUNOLOGY
Resource
Human and Animal Cells P3-X63-Ag8.653(RCB0146)