Human CD154 ELISA Kit | CD40LG elisa kit

Human CD154 ELISA Kit

Cat. Num. AAA8244625

• Gene Names: CD40LG; IGM; IMD3; TRAP; gp39; CD154; CD40L; HIGM1; T-BAM; TNFSF5; hCD40L
• Reactivity: Human
• Applications: ELISA
• Synonyms: CD154; Human CD154 ELISA Kit; CD40L; TNFSF5; TRAP; CD40 ligand; CD40-L; T-cell antigen Gp39; TNF-related activation protein; Tumor necrosis factor ligand superfamily member 5; CD40LG elisa kit

• Reactivity: Human
• Specificity: The Human CD154 ELISA Kit allows for the detection and quantification of endogenous levels of natural and/or recombinant Human CD154 proteins within the range of 156.3 pg/ml - 10000 pg/ml.
• Sequence Length: 261

• Applicable Applications for CD40LG elisa kit: Sandwich ELISA (EIA)
• Samples: Cell Culture Supernatants, Serum, Plasma.
• Sensitivity: 40 pg/ml.
• Preparation and Storage: Shipped and store at 4 degree C for 6 months, store at -20 degree C for one year. Avoid freeze/thaw cycles.

Typical Testing Data/Standard Curve

Related Product Information for CD40LG elisa kit

Principle of the Assay||The Human CD154 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human CD154 in Cell Culture Supernatants, Serum, Plasma. This assay employs an antibody specific for Human CD154 coated on a 96-well plate. Standards and samples are pipetted into the wells and CD154 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human CD154 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of CD154 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

Background/Introduction: CD40 Ligand (CD40L), also known as CD154, gp39, TNFSF5, TRAP (TNF-Related Activation Protein) or TBAM (T-cell B-cell Activating Molecule), is a multifunctional ligand in the TNF superfamily (1-4). Interaction between CD40 and CD40L is critical to the control of thymus-dependent humoral immunity and cell-mediated immune responses (5-10). The major component of the contact-dependent signal leading to B cell activation is CD40L. CD40L stimulates B cell secretion of immunoglobulin isotypes in the presence of cytokines. CD40L is a 39 kDa, 261 amino acid (aa) glycoprotein that can form homotrimers typical of other TNFSF members (1-4, 11, 12). Proteolytic cleavage can also produce 15-18 kDa soluble forms of CD40L (13, 14). Activated T cells and platelets express both a membrane-associated and a soluble form of CD40L (sCD40L) (13, 15, 16). Platelet activation during plasma and serum sample preparation can result in artificially elevated sCD40L levels (17-20). Conversely, serum samples stored above 2-8 degree C show a progressive loss of the sCD40L signal (21). sCD40L lacks the transmembrane region and a portion of the extracellular domain but contains the entire TNF- homology region. Both the membrane-bound and soluble forms of CD40L are active (22). The receptor for CD40L is CD40, a member of the TNF receptor superfamily (TNFRSF5). Interaction of CD40L with CD40 not only induces proliferation and isotype switching in B lymphocytes but also mediates a broad variety of other immune and inflammatory responses (5-7). CD40 signaling has been linked with pathogenic processes of chronic inflammatory diseases such as autoimmune diseases, neurodegenerative disorders, graft-versus-host disease, cancer, and atherosclerosis (8). The loss of interaction between CD40 and CD40L can result in impairment of T lymphocyte function, B lymphocyte differentiation, and monocyte function. CD40L is expressed primarily on activated CD4+ T cells; however, vascular endothelial cells, smooth muscle cells, macrophages, basophils, eosinophils, monocytes, dendritic cells, fibroblasts, and mast cells also express CD40L. Cytokine stimulation (e.g. IL-1beta, TNF-alpha, or IFN-gamma) can increase surface levels and de novosynthesis of CD40L in certain cell types (23). Hyper-IgM syndrome (HIGM) is an immunodeficiency characterized by elevated concentrations of serum IgM and the absence of serum IgG, IgA, and IgE. It is caused by mutations within the CD40L gene leading to defective expression on the membrane of activated T lymphocytes (24, 25). B lymphocytes from HIGM patients express functional CD40 and respond normally to wild-type CD40L, but their Tlymphocytes are unable to stimulate CD40 signaling pathways (26, 27). CD40L may play multiple roles in HIV infection (28). It may contribute to viral replication control by inducing HIV-suppressive chemokines, by downregulating monocyte cell surface expression of CCR5 and CD4, and by supporting the production of anti-HIV antibodies and cytotoxic T cells (28-31). It can also promote HIV replication in CD4+ T lymphocytes by activating antigen-presenting cells, subsequently leading to increased CD4+ T cell activation (28). With the onset of AIDS, CD40L-expressing CD4+ T cells become selectively depleted. This loss may explain the similarity between the opportunistic infections characteristic of AIDS and those observed with congenital CD40L deficiency (28). Elevated levels of sCD40L have been observed in sera from patients with systemic lupus erythematosus (SLE), chronic lymphocytic leukemia (CLL), and unstable angina (32-34). A direct relationship has been seen between disease severity and sCD40L in SLE patient sera (32). Aberrant expression of CD40L may thus contribute to autoantibody secretion in SLE through activation of bystander B lymphocytes, including cells that have been exposed to self antigens (32). Prolonged survival of malignant CLL cells may be linked to elevated levels of biologically active sCD40L (33). CD40L can mediate the resistance of CLL cells to apoptosis by Fas Ligand and fludarabine (33). Enhanced levels of both soluble and membrane-bound forms of CD40L in angina patients suggests that the CD40L-CD40 interaction may play a pathogenic role in the atherosclerotic process and in promoting acute coronary syndromes (34).

NCBI and Uniprot Product Information

• NCBI GI #: 231718
• NCBI GeneID: 959
• UniProt Accession #: P29965
• Molecular Weight: 29,274 Da
• NCBI Official Full Name: CD40 ligand
• NCBI Official Synonym Full Names: CD40 ligand
• NCBI Official Symbol: CD40LG
• NCBI Official Synonym Symbols: IGM; IMD3; TRAP; gp39; CD154; CD40L; HIGM1; T-BAM; TNFSF5; hCD40L
• NCBI Protein Information: CD40 ligand
• UniProt Protein Name: CD40 ligand
• Protein Family: CD40 ligand
• UniProt Gene Name: CD40LG
• UniProt Synonym Gene Names: CD40L; TNFSF5; TRAP; CD40-L; TRAP
• UniProt Entry Name: CD40L_HUMAN

NCBI Description

The protein encoded by this gene is expressed on the surface of T cells. It regulates B cell function by engaging CD40 on the B cell surface. A defect in this gene results in an inability to undergo immunoglobulin class switch and is associated with hyper-IgM syndrome. [provided by RefSeq, Jul 2008]

Uniprot Description

CD40LG: Mediates B-cell proliferation in the absence of co- stimulus as well as IgE production in the presence of IL-4. Involved in immunoglobulin class switching. Defects in CD40LG are the cause of X-linked immunodeficiency with hyper-IgM type 1 (HIGM1); also known as X-linked hyper IgM syndrome (XHIM). HIGM1 is an immunoglobulin isotype switch defect characterized by elevated concentrations of serum IgM and decreased amounts of all other isotypes. Affected males present at an early age (usually within the first year of life) recurrent bacterial and opportunistic infections, including Pneumocystis carinii pneumonia and intractable diarrhea due to cryptosporidium infection. Despite substitution treatment with intravenous immunoglobulin, the overall prognosis is rather poor, with a death rate of about 10% before adolescence. Belongs to the tumor necrosis factor family.

Protein type: Cell adhesion; Membrane protein, integral

Chromosomal Location of Human Ortholog: Xq26

Cellular Component: external side of plasma membrane; extracellular space; integral to membrane; integral to plasma membrane; plasma membrane

Molecular Function: CD40 receptor binding; cytokine activity; tumor necrosis factor receptor binding

Biological Process: B cell differentiation; B cell proliferation; immunoglobulin secretion; inflammatory response; isotype switching; leukocyte adhesion; negative regulation of apoptosis; platelet activation; positive regulation of interleukin-12 production; regulation of immune response; regulation of immunoglobulin secretion; tumor necrosis factor-mediated signaling pathway

Disease: Immunodeficiency With Hyper-igm, Type 1