Human CD40 Ligand/TNFSF5 Quantikine ELISA Kit

  (35 citations)     
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Assay Procedure
Citations (35)
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  • 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
QC22 , Quantikine Immunoassay Control Group 7 - Please Inquire
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.

Precision
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
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 430 1212 2638 437 1205 2612
Standard Deviation 21.8 54.4 143 27.9 72.7 162
CV% 5.1 4.5 5.4 6.4 6 6.2

Recovery

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
Linearity
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.
 CD40 Ligand/TNFSF5 [HRP]
Product Datasheets

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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; CD40L; 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.
Citations:

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.

35 Citations: Showing 1 - 10
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Species
Sample Type
  1. Influence of ethnic background on left atrial markers of inflammation, endothelial function and tissue remodelling
    Authors: CD Ruediger, B John, S Kumar, HS Lim, G Rangnekar, KC Roberts-Th, GD Young, D Chase, P Sanders, SR Willoughby
    Indian Pacing Electrophysiol J, 2018;18(1):1-5.
    Species: Human
    Sample Type: Serum
  2. Lipoprotein apheresis in the treatment of dyslipidaemia - the Czech Republic experience
    Authors: V Bláha, M Bláha, M Lánská, D Solichová, L Kujovská K, E Havel, P Vyroubal, Z Zadák, P Žák, L Sobotka
    Physiol Res, 2017;66(0):S91-S100.
    Species: Human
    Sample Type: Serum
  3. Endotoxemia contributes to CD27+ memory B-cell apoptosis via enhanced sensitivity to Fas ligation in patients with Cirrhosis
    Sci Rep, 2016;6(0):36862.
    Species: Human
    Sample Type: Plasma
  4. Potential predictive role of chemotherapy-induced changes of soluble CD40 ligand in untreated advanced pancreatic ductal adenocarcinoma
    Onco Targets Ther, 2016;9(0):4681-6.
    Species: Human
    Sample Type: Serum
  5. 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
  6. Chlorogenic acid inhibits human platelet activation and thrombus formation.
    Authors: Fuentes E, Caballero J, Alarcon M, Rojas A, Palomo I
    PLoS ONE, 2014;9(3):e90699.
    Species: Human
    Sample Type: Cell Culture Supernates
  7. The evolution of cellular deficiency in GATA2 mutation.
    Authors: Dickinson R, Milne P, Jardine L, Zandi S, Swierczek S, McGovern N, Cookson S, Ferozepurwalla Z, Langridge A, Pagan S, Gennery A, Heiskanen-Kosma T, Hamalainen S, Seppanen M, Helbert M, Tholouli E, Gambineri E, Reykdal S, Gottfreethsson M, Thaventhiran J, Morris E, Hirschfield G, Richter A, Jolles S, Bacon C, Hambleton S, Haniffa M, Bryceson Y, Allen C, Prchal J, Dick J, Bigley V, Collin M
    Blood, 2014;123(6):863-74.
    Species: Human
    Sample Type: Serum
  8. Experimental prestorage filtration removes antibodies and decreases lipids in RBC supernatants mitigating TRALI in vivo.
    Authors: Silliman C, Kelher M, Khan S, LaSarre M, West F, Land K, Mish B, Ceriano L, Sowemimo-Coker S
    Blood, 2014;123(22):3488-95.
    Species: Human
    Sample Type: Serum
  9. Involvement of Rac in thromboxane A2induced human platelet activation: regulation of sCD40 ligand release and PDGFAB secretion.
    Authors: Kageyama Y, Doi T, Matsushima-Nishiwaki R, Iida Y, Akamatsu S, Kondo A, Kuroyanagi G, Yamamoto N, Mizutani J, Otsuka T, Tokuda H, Iida H, Kozawa O, Ogura S
    Mol Med Rep, 2014;10(1):107-12.
    Species: Human
    Sample Type: Plasma
  10. 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: Platelets
  11. 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
  12. 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
  13. 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
  14. 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
  15. 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
  16. 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
  17. 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
  18. 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
  19. 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
  20. 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
  21. 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
  22. 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
  23. 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
  24. 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
  25. 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
  26. 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
  27. 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
  28. 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
  29. 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
  30. 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
  31. 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
  32. 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
  33. 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
  34. 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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