Human LIGHT/TNFSF14 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY664
Ancillary Products Available
Human LIGHT / TNFSF14 ELISA Standard Curve
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Product Details
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Citations (3)
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Human LIGHT/TNFSF14 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
62.5 - 4,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human LIGHT/TNFSF14. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

Human LIGHT / TNFSF14 ELISA Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human LIGHT/TNFSF14 DuoSet ELISA

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.

3 Citations: Showing 1 - 3
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  1. CXCL9, CXCL10, and CXCL11; biomarkers of pulmonary inflammation associated with autoimmunity in patients with collagen vascular diseases-associated interstitial lung disease and interstitial pneumonia with autoimmune features
    Authors: M Kameda, M Otsuka, H Chiba, K Kuronuma, T Hasegawa, H Takahashi, H Takahashi
    PLoS ONE, 2020-11-02;15(11):e0241719.
    Species: Human
    Sample Types: Serum
  2. The Influence of Hypertensive Therapies on Circulating Factors: Clinical Implications for SCFAs, FGF21, TNFSF14 and TNF-&alpha
    Authors: AL Magno, LY Herat, MG Kiuchi, MP Schlaich, NC Ward, VB Matthews
    Journal of Clinical Medicine, 2020-08-26;9(9):.
    Species: Human
    Sample Types: Serum
  3. Activation of platelets by Aspergillus fumigatus and potential role of platelets in the immunopathogenesis of Aspergillosis.
    Authors: Rodland EK, Ueland T, Pedersen TM, Halvorsen B, Muller F, Aukrust P, Froland SS
    Infect. Immun., 2009-12-14;78(3):1269-75.
    Species: Human
    Sample Types: Cell Culture Supernates

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