Human LIGHT/TNFSF14 Quantikine ELISA Kit

  (10 citations)     
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Assay Procedure
Citations (10)
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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 (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Saliva (25 uL)
  • Sensitivity
    16.5 pg/mL
  • Assay Range
    31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva)
  • Specificity
    Natural and recombinant human LIGHT
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
  • Interference
    No significant interference observed with available related molecules.
Product Summary
The Quantikine Human LIGHT Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human LIGHT in cell culture supernates, serum, plasma, and saliva. It contains NS0-expressed recombinant human LIGHT and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human LIGHT 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 LIGHT.

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, Saliva
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 308 626 1181 318 616 1177
Standard Deviation 11.7 25.3 34.6 27 44.2 81.8
CV% 3.8 4 2.9 8.5 7.2 6.9

Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 350 712 1388 387 754 1469
Standard Deviation 12 31.7 63.3 19.6 31.8 63.4
CV% 3.4 4.5 4.6 5.1 4.2 4.3

Recovery

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

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 105 100-113
EDTA Plasma (n=5) 106 91-114
Heparin Plasma (n=5) 101 89-115
Serum (n=5) 102 85-114
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of LIGHT were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
 LIGHT/TNFSF14 [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: LIGHT/TNFSF14
Human LIGHT is a type II membrane protein that is a member of the TNF superfamily (TNFSF14). LIGHT is an acronym that stands for is homologous to Lymphotoxins, exhibits Inducible expression, and competes with HSV Glycoprotein D for HVEM, a receptor expressed by Tlymphocytes. LIGHT has also been called HVEM-L and LT-gamma.
  • Long Name:
    TNF Ligand Superfamily Member 14
  • Entrez Gene IDs:
    8740 (Human); 50930 (Mouse)
  • Alternate Names:
    CD258 antigen; CD258; delta transmembrane LIGHT; Herpes virus entry mediator ligand; Herpesvirus entry mediator ligand; HVEM-L; HVEMLherpesvirus entry mediator-ligand; ligand for herpesvirus entry mediator; LIGHT; LIGHTherpesvirus entry mediator A; LTg; TNFSF14; TR2; tumor necrosis factor (ligand) superfamily, member 14; tumor necrosis factor ligand superfamily member 14; tumor necrosis factor receptor-like 2; tumor necrosis factor superfamily member LIGHT
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. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Additions must be completed within 15 minutes. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  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.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well.
  13.   For Cell Culture Supernate & Saliva Samples: Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.
    For Serum & Plasma Samples: Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  14. 50 µL Stop Solution
  15. 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.

10 Citations: Showing 1 - 10
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Species
Sample Type
  1. Adenovirus-mediated intratumoral expression of immunostimulatory proteins in combination with systemic Treg inactivation induces tumor-destructive immune responses in mouse models.
    Authors: Liu Y, Tuve S, Persson J, Beyer I, Yumul R, Li ZY, Tragoolpua K, Hellstrom KE, Roffler S, Lieber A
    Cancer Gene Ther., 2011;18(6):407-18.
    Species: Human
    Sample Type: Cell Culture Supernates
  2. Persistently raised plasma levels of platelet-derived inflammatory mediators in HIV-infected patients during highly active anti-retroviral therapy.
    Authors: Landro L, Ueland T, Otterdal K, Froland SS, Aukrust P
    J. Thromb. Haemost., 2011;9(5):1075-7.
    Species: Human
    Sample Type: Platelet-Poor Plasma
  3. LIGHT is associated with hypertriglyceridemia in obese subjects and increased cytokine secretion from cultured human adipocytes.
    Authors: Bassols J, Moreno-Navarrete JM, Ortega F
    Int J Obes (Lond), 2010;34(1):146-56.
    Species: Human
    Sample Type: Serum
  4. Interleukin-32 promotes osteoclast differentiation but not osteoclast activation.
    Authors: Mabilleau G, Sabokbar A
    PLoS ONE, 2009;4(1):e4173.
    Species: Human
    Sample Type: Cell Culture Supernates
  5. Inflammatory interaction between LIGHT and proteinase-activated receptor-2 in endothelial cells: potential role in atherogenesis.
    Authors: Sandberg WJ, Halvorsen B, Yndestad A, Smith C, Otterdal K, Brosstad FR, Froland SS, Olofsson PS, Damas JK, Gullestad L, Hansson GK, Oie E, Aukrust P
    Circ. Res., 2009;104(1):60-8.
    Species: Human
    Sample Type: Cell Culture Supernates
  6. Enhanced plasma levels of LIGHT in patients with acute atherothrombotic stroke.
    Authors: Liu GZ, Fang LB, Hjelmstrom P, Gao XG
    Acta Neurol. Scand., 2008;118(4):256-9.
    Species: Human
    Sample Type: Plasma
  7. The lymphotoxin-beta receptor is an upstream activator of NF-kappaB-mediated transcription in melanoma cells.
    Authors: Dhawan P, Su Y, Thu YM, Yu Y, Baugher P, Ellis DL, Sobolik-Delmaire T, Kelley M, Cheung TC, Ware CF, Richmond A
    J. Biol. Chem., 2008;283(22):15399-408.
    Species: Human
    Sample Type: Cell Culture Supernates
  8. Platelet-associated LIGHT (TNFSF14) mediates adhesion of platelets to human vascular endothelium.
    Authors: Celik S, Langer H, Stellos K, May AE, Shankar V, Kurz K, Katus HA, Gawaz MP, Dengler TJ
    Thromb. Haemost., 2007;98(4):798-805.
    Species: Human
    Sample Type: Cell Culture Supernates
  9. LIGHT (TNFSF14), a novel mediator of bone resorption, is elevated in rheumatoid arthritis.
    Authors: Edwards JR, Sun SG, Locklin R, Shipman CM, Adamopoulos IE, Athanasou NA, Sabokbar A
    Arthritis Rheum., 2006;54(5):1451-62.
    Species: Human
    Sample Type: Serum
  10. Platelet-derived LIGHT induces inflammatory responses in endothelial cells and monocytes.
    Authors: Otterdal K, Smith C, Oie E, Pedersen TM, Yndestad A, Stang E, Endresen K, Solum NO, Aukrust P, Damas JK
    Blood, 2006;108(3):928-35.
    Species: Human
    Sample Type: Plasma

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