Human sCD40L ELISA Kit (96-well)

CAT#: EA100114

Human sCD40L ELISA Kit (96-well)

Product Images

Representative standard curve for sCD40L ELISA. sCD40L was diluted in serial two-fold steps in Sample Diluent.

Specifications

Product Data

• Format: 96-well strip plate
• Assay Type: Solid Phase Sandwich ELISA
• Assay Length: 3 hours
• Signal: Colorimetric
• Curve Range: 156.25-10000pg/ml
• Sample Type: Cell culture supernatant, serum, plasma (EDTA, citrate, heparin)
• Sample Volume: 20 uL
• Specificity: Natural and recombinant Human sCD40L Ligand
• Sensitivity: 40pg/mL
• Reactivities: Human
• Interference: No significant interference observed with available related molecules.
• Components:
  • Aluminium pouches with a Microwell Plate coated with antibody to human sCD40L (8X12)
  • 2 vials human sCD40L Standard lyophilized, 10000 pg/ml upon reconstitution
  • 2 vials HRP-Conjugate solution
  • 1 bottle Standard /sample Diluent
  • 1 bottle Streptavidin-HRP Diluent
  • 1 bottle Wash Buffer Concentrate 20x (PBS with 1% Tween-20)
  • 1 vial Substrate Solution
  • 1 vial Stop Solution
  • 4 pieces Adhesive Films
  • package insert
• Background:

  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 . Interaction between CD40 and CD40L is critical to the control of thymus-dependent humoral immunity and cell-mediated immune responses . 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 . Proteolytic cleavage can also produce 15-18 kDa soluble forms of CD40L . Activated T cells and platelets express both a membrane-associated and a soluble form of CD40L (sCD40L) . Platelet activation during plasma and serum sample preparation can result in artificially elevated sCD40L levels . Conversely, serum samples stored above 2-8 °C show a progressive loss of the sCD40L signal . 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 .

  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 . 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 . 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-1., TNF-., or IFN-.) can increase surface levels and de novo synthesis of CD40L in certain cell types . 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 . B lymphocytes from HIGM patients express functional CD40 and respond normally to wild-type CD40L, but their T lymphocytes are unable to stimulate CD40 signaling pathways .

  CD40L may play multiple roles in HIV infection . 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 . It can also promote HIV replication in CD4+ T lymphocytes by activating antigen-presenting cells, subsequently leading to increased CD4+ T cell activation . 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 .

  Elevated levels of sCD40L have been observed in sera from patients with systemic lupus erythematosus (SLE), chronic lymphocytic leukemia (CLL), and unstable angina . A direct relationship has been seen between disease severity and sCD40L in SLE patient sera . 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 . Prolonged survival of malignant CLL cells may be linked to elevated levels of biologically active sCD40L . CD40L can mediate the resistance of CLL cells to apoptosis by Fas Ligand and fludarabine . Enhanced levels of both

• Gene Symbol: CD40LG
• Standard Curve:
  
  Representative standard curve for sCD40L ELISA. sCD40L was diluted in serial two-fold steps in Sample Diluent.