Human CD40 Ligand/TNFSF5 Quantikine ELISA Kit

(27 citations)   
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (100 uL), Serum (20 uL), Platelet-poor EDTA Plasma (100 uL), Platelet-poor Heparin Plasma (100 uL)
  • Sensitivity
    10.1 pg/mL
  • Assay Range
    62.5 - 4,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma)
  • Specificity
    Natural and recombinant human CD40 Ligand
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
Product Summary
The Quantikine Human CD40 Ligand Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human CD40 Ligand in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human CD40 Ligand and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human CD40 Ligand showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human CD40 Ligand.

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Standard Deviation21.854.414327.972.7162


The recovery of CD40 Ligand spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 102 90-114
Platelet-poor EDTA Plasma (n=5) 98 86-107
Platelet-poor Heparin Plasma (n=5) 94 86-113
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of CD40 Ligand were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human CD40 Ligand/TNFSF5 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: CD40 Ligand/TNFSF5
CD40 Ligand, also known as TNFSF5, CD154, TRAP, and gp39, is a trimeric protein that is expressed in transmembrane and soluble forms. It is expressed by many immune cell types as well as by endothelial and smooth muscle cells of the vasculature. It binds to CD40 on antigen presenting cells, platelets, and endothelial and epithelial cells, leading to enhanced B cell activation and T cell dependent humoral immune responses. CD40 Ligand dysregulation contributes to immune pathology in AIDS, atherosclerosis, atherothrombosis, and restenosis.
    • Entrez Gene IDs
      959 (Human); 21947 (Mouse); 84349 (Rat);
    • Alternate Names
      CD154 antigen; CD154; CD40 antigen ligand; CD40 ligand; CD40-L; CD40LG; CD40LIGM; gp39; hCD40L; HIGM1; T-B cell-activating molecule; T-BAM; T-cell antigen Gp39; TNF-related activation protein; TNFSF5; TNFSF5IMD3; TRAP; TRAPtumor necrosis factor (ligand) superfamily, member 5 (hyper-IgM syndrome); tumor necrosis factor (ligand) superfamily member 5; Tumor necrosis factor ligand superfamily member 5;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 100 µL Assay Diluent
    4.   Add 100 µL of Assay Diluent to each well.

    5. 100 µL Standard, Control, or Sample
    6.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
    10.   Aspirate and wash 4 times.

    11. 200 µL Substrate Solution
    12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

    13. 50 µL Stop Solution
    14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

    R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

    Showing Results 1 - 10 of 27
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    Sample Type
    1. Inhibition of platelet activation and thrombus formation by adenosine and inosine: studies on their relative contribution and molecular modeling.
      Authors: Fuentes E, Pereira J, Mezzano D, Alarcon M, Caballero J, Palomo I
      PLoS ONE, 2014;9(11):e112741.
      Species: Human
      Sample Type: Cell Culture Supernates
    2. Protective mechanisms of guanosine from Solanum lycopersicum on agonist-induced platelet activation: role of sCD40L.
      Authors: Fuentes E, Alarcon M, Astudillo L, Valenzuela C, Gutierrez M, Palomo I
      Molecules, 2013;18(7):8120-35.
      Species: Human
      Sample Type: Platelet Concentrate
    3. Elderly trauma patients have high circulating noradrenaline levels but attenuated release of adrenaline, platelets, and leukocytes in response to increasing injury severity.
      Authors: Johansson PI, Sorensen AM, Perner A, Welling KL, Wanscher M, Larsen CF, Ostrowski SR
      Crit. Care Med., 2012;40(6):1844-50.
      Species: Human
      Sample Type: Plasma
    4. Decreased plasma cytokines are associated with low platelet counts in aplastic anemia and immune thrombocytopenic purpura.
      J. Thromb. Haemost., 2012;10(8):1616-23.
      Species: Human
      Sample Type: Serum
    5. Prospective study of circulating soluble CD40 ligand concentrations and the incidence of cardiovascular disease in a nested prospective case-control study of older men and women.
      Authors: Jefferis BJ, Whincup PH, Welsh P, Wannamethee SG, Rumley A, Lawlor DA, Ebrahim S, Lowe GD
      J. Thromb. Haemost., 2011;9(8):1452-9.
      Species: Human
      Sample Type: Serum
    6. High sCD40L levels early after trauma are associated with enhanced shock, sympathoadrenal activation, tissue and endothelial damage, coagulopathy and mortality.
      Authors: Johansson PI, Sorensen AM, Perner A, Welling KL, Wanscher M, Larsen CF, Ostrowski SR
      J. Thromb. Haemost., 2011;10(2):207-16.
      Species: Human
      Sample Type: Plasma
    7. Matrix metalloproteinase-9 and plasminogen activator inhibitor-1 are associated with right ventricular structure and function: the MESA-RV Study.
      Authors: Kawut SM, Barr RG, Johnson WC
      Biomarkers, 2010;15(8):731-8.
      Species: Human
      Sample Type: Plasma
    8. Pre-operative platelet-bound CD40 ligand is probably associated with peri-operative cardiac events in hip and knee arthroplasty.
      Authors: Ray MJ, Calabro LJ, Sirisena T, Crawford SA, Crawford RW, Walters DL
      Eur. J. Clin. Invest., 2010;40(6):497-503.
      Species: Human
      Sample Type: Plasma
    9. A plant stanol yogurt drink alone or combined with a low-dose statin lowers serum triacylglycerol and non-HDL cholesterol in metabolic syndrome patients.
      Authors: Plat J, Brufau G, Dallinga-Thie GM, Dasselaar M, Mensink RP
      J. Nutr., 2009;139(6):1143-9.
      Species: Human
      Sample Type: Plasma
    10. Platelet turnover, coagulation factors, and soluble markers of platelet and endothelial activation in essential thrombocythemia: relationship with thrombosis occurrence and JAK2 V617F allele burden.
      Authors: Arellano-Rodrigo E, Alvarez-Larran A, Reverter JC, Colomer D, Villamor N, Bellosillo B, Cervantes F
      Am. J. Hematol., 2009;84(2):102-8.
      Species: Human
      Sample Type: Plasma
    11. Interleukin-18 is a strong predictor of cardiovascular events in elderly men with the metabolic syndrome: synergistic effect of inflammation and hyperglycemia.
      Authors: Troseid M, Seljeflot I, Hjerkinn EM, Arnesen H
      Diabetes Care, 2009;32(3):486-92.
      Species: Human
      Sample Type: Plasma
    12. Hyperglycemia is associated with enhanced thrombin formation, platelet activation, and fibrin clot resistance to lysis in patients with acute coronary syndrome.
      Authors: Undas A, Wiek I, Stepien E, Zmudka K, Tracz W
      Diabetes Care, 2008;31(8):1590-5.
      Species: Human
      Sample Type: Plasma
    13. Plasma concentration of SCUBE1, a novel platelet protein, is elevated in patients with acute coronary syndrome and ischemic stroke.
      Authors: Dai DF, Thajeb P, Tu CF, Chiang FT, Chen CH, Yang RB, Chen JJ
      J. Am. Coll. Cardiol., 2008;51(22):2173-80.
      Species: Human
      Sample Type: Plasma
    14. Fish oil in combination with high or low intakes of linoleic acid lowers plasma triacylglycerols but does not affect other cardiovascular risk markers in healthy men.
      Authors: Damsgaard CT, Frokiaer H, Andersen AD, Lauritzen L
      J. Nutr., 2008;138(6):1061-6.
      Species: Human
      Sample Type: Serum
    15. Soluble CD40 ligand as a biomarker for storage-related preanalytic variations of human serum.
      Authors: Lengelle J, Panopoulos E, Betsou F
      Cytokine, 2008;44(2):275-82.
      Species: Human
      Sample Type: Serum
    16. Analysis of platelet alpha2-adrenergic receptor activity in stable coronary artery disease patients on dual antiplatelet therapy.
      Authors: Beres BJ, Toth-Zsamboki E, Vargova K, László A, Masszi T, Kerecsen G, Preda I, Kiss RG
      Thromb. Haemost., 2008;100(5):829-38.
      Species: Human
      Sample Type: Plasma
    17. Rosiglitazone produces a greater reduction in circulating platelet activity compared with gliclazide in patients with type 2 diabetes mellitus--an effect probably mediated by direct platelet PPARgamma activation.
      Authors: Khanolkar MP, Morris RH, Thomas AW, Bolusani H, Roberts AW, Geen J, Jackson SK, Evans LM
      Atherosclerosis, 2007;197(2):718-24.
      Species: Human
      Sample Type: Serum
    18. Multiple biomarker use for detection of adverse events in patients presenting with symptoms suggestive of acute coronary syndrome.
      Authors: Apple FS, Pearce LA, Chung A, Ler R, Murakami MM
      Clin. Chem., 2007;53(5):874-81.
      Species: Human
      Sample Type: Plasma
    19. Relationship of soluble CD40 ligand to vascular endothelial growth factor, angiopoietins, and tissue factor in atrial fibrillation: a link among platelet activation, angiogenesis, and thrombosis?
      Authors: Choudhury A, Freestone B, Patel J, Lip GY
      Chest, 2007;132(6):1913-9.
      Species: Human
      Sample Type: Plasma
    20. Myeloid-related protein 8/14 complex is released by monocytes and granulocytes at the site of coronary occlusion: a novel, early, and sensitive marker of acute coronary syndromes.
      Authors: Altwegg LA, Neidhart M, Hersberger M, Muller S, Eberli FR, Corti R, Roffi M, Sutsch G, Gay S, von Eckardstein A, Wischnewsky MB, Luscher TF, Maier W
      Eur. Heart J., 2007;28(8):941-8.
      Species: Human
      Sample Type: Serum
    21. Relation of improvement in endothelium-dependent flow-mediated vasodilation after rosiglitazone to changes in asymmetric dimethylarginine, endothelin-1, and C-reactive protein in nondiabetic patients with the metabolic syndrome.
      Authors: Wang TD, Chen WJ, Cheng WC, Lin JW, Chen MF, Lee YT
      Am. J. Cardiol., 2006;98(8):1057-62.
      Species: Human
      Sample Type: Plasma
    22. No difference in the effects of clopidogrel and aspirin on inflammatory markers in patients with coronary heart disease.
      Authors: Solheim S, Pettersen AA, Arnesen H, Seljeflot I
      Thromb. Haemost., 2006;96(5):660-4.
      Species: Human
      Sample Type: Plasma
    23. Effects of simvastatin on angiogenic growth factors released at the site of microvascular injury.
      Authors: Undas A, Celinska-Lowenhoff M, Stepien E, Nizankowski R, Tracz W, Szczeklik A
      Thromb. Haemost., 2006;95(6):1045-7.
      Species: Human
      Sample Type: Plasma
    24. Humoral and cellular factors responsible for coronary collateral formation.
      Authors: Sherman JA, Hall A, Malenka DJ, De Muinck ED, Simons M
      Am. J. Cardiol., 2006;98(9):1194-7.
      Species: Human
      Sample Type: Plasma
    25. Effects of rosiglitazone alone and in combination with atorvastatin on nontraditional markers of cardiovascular disease in patients with type 2 diabetes mellitus.
      Authors: Chu CS, Lee KT, Lee MY, Su HM, Voon WC, Sheu SH, Lai WT
      Am. J. Cardiol., 2006;97(5):646-50.
      Species: Human
      Sample Type: Serum
    26. Additive beneficial effects of fenofibrate combined with candesartan in the treatment of hypertriglyceridemic hypertensive patients.
      Authors: Koh KK, Quon MJ, Han SH, Chung WJ, Ahn JY, Kim JA, Lee Y, Shin EK
      Diabetes Care, 2006;29(2):195-201.
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