Monoclonal antibodies directed against chicken ?2-microglobulin developed with a synthesized peptide.

ABSTRACT: We developed a panel of monoclonal antibodies (MAb) against chicken ?2-microglobulin (ch?2M) by fusions between SP2/0 myeloma cells and spleen cells from mice immunized with a synthesized peptide corresponding to positions 91-119 of the COOH domain of ch?2M. Two of them, 6E7 and 3D1, identified as IgG1/?, could react with ch?2M protein from avian macrophage HD11 cells and human 293T cells transfected with pcDNA3.1-ch?2M in immunofluorescence assays. Only a 12?kDa protein band of ch?2M could be detected in the HD11 and 293T/ch?2M cell lysates by Western blot analysis. Chicken ?2M in serum and plasma could be found in Western blot by MAb 3D1. Moreover, MAb 3D1 also recognized the ch?2M antigen on the cell membranes in flow cytometry. Immunohistochemical staining with these MAbs revealed that ch?2M was present in chicken thymus, spleen, and bursa. These MAbs will be good tools for analyzing the mechanism of the chicken immune system.

SUBMITTER: Yu C 

PROVIDER: S-EPMC3733132 | biostudies-literature | 2013 Jun

REPOSITORIES: biostudies-literature