Human LIGHT/TNFSF14 Quantikine ELISA Kit

R&D Systems | Catalog # DLIT00

R&D Systems
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL), Saliva (25 µL)

Sensitivity

16.5 pg/mL

Assay Range

31.2-2000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva)
View all LIGHT/TNFSF14 ELISA Kits »

Product Summary for Human LIGHT/TNFSF14 Quantikine ELISA Kit

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.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

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.

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 (pg/mL) 308 626 1181 318 616 1177
Standard Deviation 11.7 25.3 34.6 27.0 44.2 81.8
CV% 3.8 4.0 2.9 8.5 7.2 6.9

EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 350 712 1388 387 754 1469
Standard Deviation 12.0 31.7 63.3 19.6 31.8 63.4
CV% 3.4 4.5 4.6 5.1 4.2 4.3

Recovery for Human LIGHT/TNFSF14 Quantikine ELISA Kit

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

Scientific Data Images for Human LIGHT/TNFSF14 Quantikine ELISA Kit

Human LIGHT/TNFSF14 ELISA Calibrator Diluent RD5K Standard Curve

Human LIGHT/TNFSF14 ELISA Calibrator Diluent RD5K Standard Curve

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

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

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

Alternate Names

CD258, TNFSF14

Entrez Gene IDs

8740 (Human); 50930 (Mouse)

Gene Symbol

TNFSF14

Additional LIGHT/TNFSF14 Products

Product Documents for Human LIGHT/TNFSF14 Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human LIGHT/TNFSF14 Quantikine ELISA Kit

For research use only

⚠ 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.

Citations for Human LIGHT/TNFSF14 Quantikine ELISA Kit

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    Sample Tested: 3A-sub E human placenta cell line
    Verified Customer | Posted 06/04/2019

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Protocols

View specific protocols for Human LIGHT/TNFSF14 Quantikine ELISA Kit (DLIT00):

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
  3.   Add 100 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   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.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well.
  9.   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.
    10. 50 µL Stop Solution
  10. 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.

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