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  • Development and characterisation of monoclonal antibodies reactive with porcine CSF1R (CD115).

Development and characterisation of monoclonal antibodies reactive with porcine CSF1R (CD115).

Developmental and comparative immunology (2014-07-16)
L Moffat, L Rothwell, C Garcia-Morales, K A Sauter, R Kapetanovic, D J Gow, D A Hume
ABSTRACT

Macrophage colony-stimulating factor (CSF1) controls the proliferation and differentiation of cells of the mononuclear phagocyte system. CSF1, alongside a second ligand, interleukin-34 (IL-34), acts by binding to a cell surface receptor (CSF1R). We previously cloned and expressed pig CSF1 and IL-34. Here we produced a pig CSF1R-Ig+pFUSE Fc fusion protein and used it as an immunogen to produce three monoclonal antibodies (ROS8G11, ROS3A5 and ROS3B10) targeted against porcine CSF1R. Specific binding of each monoclonal antibody was confirmed by ELISA, Western blot, flow cytometry and immunocytochemistry. The antibodies did not block CSF1 signalling. The surface expression of CSF1R in pig peripheral blood was restricted to CD14-positive monocytes and was also detected on lung macrophages. These antibodies provided an opportunity to investigate the increase of available CSF1R during pig BMDM differentiation. The new monoclonal antibodies provide useful reagents to support the study of monocyte and macrophage biology in the pig.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
IL-6 from mouse, Animal-component free, recombinant, expressed in E. coli, ≥98% (SDS-PAGE), ≥98% (HPLC)
Sigma-Aldrich
Interleukin-6 from mouse, IL-6, recombinant, expressed in E. coli, lyophilized powder, suitable for cell culture, carrier free
Sigma-Aldrich
DAPI, for nucleic acid staining