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Human LIGHT/TNFSF14

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 CD40 / TNFRSF5

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.

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.

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

Assay Procedure

Refer to the product datasheet for the 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.

1Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3Add 50 μL of Assay Diluent to each well.

4Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7Aspirate and wash 5 times.

8Add 100 μL Substrate Solution to each well.

9Add 100 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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