CD40L ELISA Kit (Human) (OKBB00282)

Catalog No: OKBB00282

Size:96 Tests

Human CD40LG ELISA Kit standard curve

Product Info

• Predicted Species Reactivity: Human
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's human CD40L ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for CD40L has been precoated onto 96-well plates. Standards(E.coli, E108-L261) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for CD40L is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The absorbance of yellow is proportional to the human CD40L amount of sample captured in plate.
• ELISA Kit Range: 62.5 pg/ml - 4,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (ng/mL)	0.44	1.26	2.38	0.42	1.41	2.46
  St. Dev.	0.026	0.064	0.145	0.029	0.086	0.182
  %CV	5.90	5.07	6.09	6.90	6.09	7.39

• ELISA Kit Component:
  • Lyophilized recombinant human CD40LG standard: 10 ng/tube x 2
  • Anti-human CD40LG Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-human CD40LG antibody: 130 uL, dilution 1:100
  • Antibody diluent buffer: 12 mL
  • Avidin-Biotin-Peroxidase Complex (ABC): 130 uL, dilution 1:100
  • ABC diluent buffer: 12 mL
  • TMB color developing agent: 10 mL
  • TMB stop solution: 10 mL
• Additional Information: Range: 62.5pg/ml-4000pg/ml
  Principle: Aviva’s human CD40L ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human CD40L specific-specific polyclonal antibodies were precoated onto 96-well plates. The human specific detection polyclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human CD40L amount of sample captured in plate.
  Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot
  experiment using standards and a small number of samples is recommended.
  2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
  3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
  4. Duplicate well assay is recommended for both standard and sample testing.
  5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
  6. Don’t reuse tips and tubes to avoid cross contamination.
  7. To avoid to use the reagents from different batches together.
  8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the
  marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before
  using.
  Background: CD40 ligand(CD40L) is a type II membrane protein of 261 amino acids on activated T cells that induces B cell proliferation and immunoglobulin secretion. It has homology with tumour necrosis factor-alpha and -beta, and has important functions in B-cell activation and differentiation. Human CD40L with 5 exons, is mapped to the proximal region of the mouse X chromosome on Xq26.3-27.1, and can be detected on T cells but is absent from B cells and monocytes. Since CD40L is expressed on platelets and released from them on activation, its predictive value as a marker for clinical outcome and the therapeutic effect of inhibition of glycoprotein IIb /IIIa receptor in patients with acute coronary syndromes was investigated. The soluble CD40L may be involved in the process of restenosis and that it exerts its effect by triggering a complex group of inflammatory reactions on endothelial and mononuclear cells.CD40L plays a central role in the pathophysiology of acute coronary syndromes, and has a role in the pathogenesis of coronary artery lesions.
• Reconstitution and Storage: Store at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
• Sample Type: cell culture supernates, serum and plasma (heparin, EDTA)
• Sensitivity: <15 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine
• Reference: 1. Allen,R.C.;Armitage,R.J.;Conley,M.E.;Rosenblatt,H.;Jenkins,N.A.;Copeland,N.G.;Bedell,M.A.;Edelhoff,S.;Disteche,C.
  M.;Simoneaux,D.K.;Fanslow,W.C.;Belmont,J.;Spriggs,M.K.:CD40 ligand gene defects responsible for X-linked
  hyper-IgM syndrome. Science 259: 990-993, 1993.
  2. Cipollone,F.;Ferri,C.;Desideri,G.;Paloscia,L.;Materazzo,G.;Mascellanti,M.;Fazia,M.;Iezzi,A.;Cuccurullo,C.;Pini,B.;Buc
  ci,M.;Santucci,A.;Cuccurullo,F.;Mezzetti,A.:Preprocedural level of soluble CD40L is predictive of enhanced
  inflammatory response and restenosis after coronary angioplasty. Circulation 108: 2776-2782, 2003.
• Specificity: Natural and recombinant human CD40L
• Immunogen: Expression system for standard: E.coli; Immunogen sequence: E108-L261
• Description: For quantitative detection of human soluble CD40L in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Target Info

• Gene Symbol: CD40LG
• Gene Full Name: CD40 ligand
• Alias Symbols: CD154, CD40 antigen ligand, CD40 ligand, CD40-L, CD40L_HUMAN, CD40LG, gp39, HIGM1, IGM, IMD3, soluble form, T B cell activating molecule, T BAM, T cell antigen Gp39, T-cell antigen Gp39, TNF related activation protein, TNF-related activation protein, TNFSF5, TrAP, Tumor necrosis factor (ligand) superfamily member 5, Tumor necrosis factor (ligand) superfamily, member 5 (hyper-IgM syndrome), Tumor necrosis factor ligand superfamily member 5
• NCBI Gene Id: 959
• Protein Name: CD40 ligand
• Description of Target: CD40 ligand(CD40L) is a type II membrane protein of 261 amino acids on activated T cells that induces B cell proliferation and immunoglobulin secretion. It has homology with tumour necrosis factor-alpha and -beta, and has important functions in B-cell activation and differentiation. Human CD40L with 5 exons, is mapped to the proximal region of the mouse X chromosome on Xq26.3-27.1, and can be detected on T cells but is absent from B cells and monocytes. Since CD40L is expressed on platelets and released from them on activation, its predictive value as a marker for clinical outcome and the therapeutic effect of inhibition of glycoprotein IIb /IIIa receptor in patients with acute coronary syndromes was investigated. The soluble CD40L may be involved in the process of restenosis and that it exerts its effect by triggering a complex group of inflammatory reactions on endothelial and mononuclear cells.CD40L plays a central role in the pathophysiology of acute coronary syndromes, and has a role in the pathogenesis of coronary artery lesions.
• Uniprot ID: P29965
• Protein Accession #: NP_000065.1
• Nucleotide Accession #: NM_000074.2