Human LIGHT/TNFSF14 Quantikine ELISA Kit

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DLIT00
Human LIGHT/TNFSF14 ELISA Linearity
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Citations (10)
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Human LIGHT/TNFSF14 Quantikine ELISA Kit Summary

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

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

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

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.
Human LIGHT/TNFSF14 ELISA Linearity

Data Examples

Human LIGHT/TNFSF14 ELISA Calibrator Diluent RD5K Standard Curve

Human LIGHT/TNFSF14 ELISA Calibrator Diluent RD6-6 Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: LIGHT/TNFSF14

LIGHT, also known as TNFSF14, is a homotrimeric glycoprotein in the TNF superfamily. It is expressed on the surface of activated lymphocytes, NK cells, immature dendritic cells, monocytes, and granulocytes. A soluble form can be shed by proteolysis. LIGHT binds and activates HVEM/TNFRSF14 to costimulate Th1 immune responses, enhance CD8+ T cell mediated tumor immunity, and regulate allogeneic T cell activation and allograft rejection. In addition, it binds and activates Lymphotoxin beta R/TNFRSF3 to induce apoptosis, cytokine production, mesenteric lymph node organogenesis, and maintenance of lymph node structure and function. It can also bind the soluble human decoy receptor DcR3/TNFRSF6B.

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
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

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 for Human LIGHT/TNFSF14 Quantikine ELISA Kit

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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  1. 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 Types: Platelet Poor Plasma
  2. 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 Types: Cell Culture Supernates
  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 Types: 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 Types: 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 Types: 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 Types: 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 Types: 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 Types: Cell Culture Supernates
  9. 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 Types: Plasma
  10. 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 Types: Serum

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Human LIGHT/TNFSF14 Quantikine ELISA Kit
By Anonymous on 06/04/2019
Sample Tested: 3A-sub E human placenta cell line