|Detection of Human CD40/TNFRSF5 by Western Blot. Western blot shows lysates of Raji human Burkitt's lymphoma cell line and Daudi human Burkitt's lymphoma cell line. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human CD40/TNFRSF5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF632) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for CD40/TNFRSF5 at approximately 40-45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
|Human CD40/TNFRSF5 Antibody Stimulates Cell Proliferation in Human B Cells. Goat Anti-Human CD40/TNFRSF5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF632) stimulates human B cell proliferation in the presence of Recombinant Human IL‑4 (Catalog # 204-IL) in a dose-dependent manner, as measured by Resazurin (Catalog # AR002). The ED50 for this effect is typically 3-12 ng/mL|
CD40 is a type I transmembrane glycoprotein belonging to the TNF receptor superfamily. The mature hCD40 consists of a 172 amino acid (aa) extracellular domain, a 22 aa transmembrane region and a 62 aa cytoplasmic domain (1). Human and mouse CD40 share 62% aa identity. CD40 is expressed in B cells, follicular dendritic cells, dendritic cells, activated monocytes, macrophages, endothelial cells, vascular smooth muscle cells, and several tumor cell lines (2). The extracellular domain has the cysteine-rich repeat regions, which are characteristic for many of the receptors of the TNF superfamily. Interaction of CD40 with its ligand, CD40L, leads to aggregation of CD40 molecules, which in turn interact with cytoplasmic components to initiate signaling pathways. Early studies on the CD40-CD40L system revealed its role in humoral immunity. Interaction between CD40L on T cells and CD40 on B cells stimulated B cell proliferation and provided the signal for immunoglobulin isotype switching (3). Mutations in the CD40L gene, which resulted in a CD40L molecule unable to interact with CD40, are responsible for the hyper-IgM syndrome (4). Cross-linking of CD40 with antibodies or by CD40 binding to CD40L produces cell type-specific responses which include costimulation and induction of proliferation, induction of cytokine production, rescue from apoptosis, and upregulation of adhesion molecules (5). Some of the early events of intracellular signaling by the CD40-CD40L system include the association of the CD40 with TRAFs and the activation of various kinases (6 - 8).
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