Human LIGHT/TNFSF14 | ||||||||||||||||||||||||||||
Human LIGHT/TNFSF14 Quantikine ELISA Kit Summary
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
Background: LIGHT/TNFSF14LIGHT, 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.
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. 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、Add 50 μL of Assay Diluent to each well. 4、Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours. 5、Aspirate each well and wash, repeating the process 4 times for a total of 5 washes. 6、Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours. 7、Aspirate and wash 5 times. 8、Add 100 μL Substrate Solution to each well. 9、Add 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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