CD40 is a 45-50-kDa glycoprotein belonging to the tumor necrosis factor (TNF) receptor superfamily. CD40 is specifically expressed on the surface of B cells and specialized antigen-presenting cells such as dendritic cells and macrophages. CD40 interacts with the CD40 ligand (CD154) which is found primarily on T cells, playing a role in both humoral and cell-mediated immune responses. Activation of CD40 on B cells by CD40 ligand causes B cell proliferation, differentiation, immunoglobulin isotype switching, germinal center formation, and stimulation of the humoral memory response.
This antibody reacts with a 45-48 kDa type I integral membrane glycoprotein present on peripheral blood and tonsillar B cells, but not expressed on terminally differentiated B cells.
The antibody against human CD40 was produced from hybridoma (5C3) cultured in serum-free medium and purified under mild conditions by proprietary chromatography processes.
Immunohistochemistry (acetone-fixed frozen section; indirect immuno-staining)
Isotype: Mouse IgG1κ
Immunogen: Recombinant extracellular domain of CD40
Conjugate: [Biotin] / [IgG] = 7.1
Form: 0.75mg/ml in PBS, 50% glycerol, filter-sterilized
Storage: Shipped at 4°C and stored at -20°C
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Fig.1 Detection of endogenous CD40 in whole cell extract of MCF7 cells by Western blot with biotin conjugated anti-CD40 antibody (5C3). Proteins in the extract were separated by 12.5% SDS-PAGE and blotted by wet system. Biotin-conjugated antibody (5C3) was used at 1/1,000 dilution and HRP-conjugated streptoavidin was used at 1/1000 dilution.
Fig. 2 Flow-cytometry analysis of human CD40 expression in transgenic mouse. Splenocytes from m (mouse) CD40+/- , mCD40-/- and hCD40 wild type/mCD40-/- mice were stained with monoclonal antibodies against mCD40, B220 and hCD40 (5C3) and analyzed by flow cytometry. hCD40 molecules were expressed specifically on B220+ B cells.