Jiro Hirota1, Shinya Shimizu1, Atsushi Watanabe2, Fumiko Suzuta3, Kazue Yajima4, Kumiko Kimura5, Makoto Haritani5, Shigeki Inumaru1, Yukio Yagi6
European Cytokine Network, Vol.22, No.1, pp. 73-80, 2011, DOI:10.1684/ecn.2011.0275
Abstract Three IgG class anti-bovine CXCL8 (bCXCL8) monoclonal antibody (mAb)-secreting hybridomas,
SH8-8D7, SH8-12A5 and SH8-2A1, were developed. SH8-8D7 was IgG2a, and SH8-12A5 and SH8-2A1 were IgG1.
All three mAbs detected recombinant bCXCL8 (rbCXCL8) by immunoprecipitation and Western blotting. SH8-
2A1 could neutralise the chemotactic activity of rbCXCL8 towards neutrophils. The quantitative bCXCL8 ELISA
was constituted by the combination of SH8-12A5 and biotin-SH8-2A1. The detection range was 20-1000 pg/mL. A
sandwich ELISA was used to measure native bCXCL8 derived from the supernatant of cultured bovine peripheral
blood mononuclear cells stimulated with ConA, LPS or PHA. Furthermore, SH8-2A1 More >