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Human DcR3/TNFRSF6B DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
187.0 - 12,000 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 DcR3/TNFRSF6B. 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)

Data Example

Human DcR3 / TNFRSF6B ELISA 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.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: DcR3/TNFRSF6B

Decoy receptor 3 (DcR3), also called TNFRSF6B, TR6 and M68, is a member of the TNF receptor superfamily. DcR3 lacks a transmembrane sequence and is a secreted protein. DcR3 shares sequence identity with OPG (31%), TNF RII (29%) and Fas (17%), and is expressed in a variety of different tissues and at high levels in many malignant tumors.

Long Name:
Decoy Receptor 3
Entrez Gene IDs:
8771 (Human); 107126607 (Cynomolgus Monkey)
Alternate Names:
DcR3; DCR3Decoy receptor for Fas ligand; Decoy receptor 3; M68; M68DJ583P15.1.1; TNFRSF6B; TR6; TR6DcR3; tumor necrosis factor receptor superfamily member 6B; tumor necrosis factor receptor superfamily, member 6b, decoy

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 DcR3/TNFRSF6B 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.

13 Citations: Showing 1 - 10
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  1. Hepatitis B X protein upregulates decoy receptor 3 expression via the PI3K/NF-?B pathway
    Authors: DY Liang, S Sha, Q Yi, J Shi, Y Chen, Y Hou, Q Chang
    Cell. Signal., 2019;62(0):109346.
    Species: Human
    Sample Types: Serum
  2. DcR3 induces proliferation, migration, invasion, and EMT in gastric cancer cells via the PI3K/AKT/GSK-3?/?-catenin signaling pathway
    Authors: H Ge, C Liang, Z Li, D An, S Ren, C Yue, J Wu
    Onco Targets Ther, 2018;11(0):4177-4187.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Downregulation of DcR3 sensitizes hepatocellular carcinoma cells to TRAIL-induced apoptosis
    Authors: C Liang, Y Xu, G Li, T Zhao, F Xia, G Li, D Zhang, J Wu
    Onco Targets Ther, 2017;10(0):417-428.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Elevated Serum Levels of the Antiapoptotic Protein Decoy-Receptor 3 Are Associated with Advanced Liver Disease
    Authors: Spiros D Ladas
    Can J Gastroenterol Hepatol, 2016;2016(0):2637010.
    Species: Human
    Sample Types: Serum
  5. Inflammatory markers and clinical characteristics for predicting persistent positivity of interferon gamma release assay in dialysis population.
    Authors: Chin-Chung Shu, Chia-Lin Hsu, Chih-Yuan Lee, Vin-Cent Wu, Feng-Jung Yang, Jann-Yuan Wang, Chong-Jen Yu, Li-Na Lee
    Scientific Reports, 2016;0(0):2045-2322.
    Species: Human
    Sample Types: Plasma
  6. Chronic lymphocytic leukemia: a paradigm of innate immune cross-tolerance.
    Authors: Jurado-Camino T, Cordoba R, Esteban-Burgos L, Hernandez-Jimenez E, Toledano V, Hernandez-Rivas J, Ruiz-Sainz E, Cobo T, Siliceo M, Perez de Diego R, Belda C, Cubillos-Zapata C, Lopez-Collazo E
    J Immunol, 2015;194(2):719-27.
    Species: Human
    Sample Types: Serum
  7. Apoptosis-associated biomarkers in tuberculosis: promising for diagnosis and prognosis prediction.
    Authors: Shu, Chin-Chu, Wu, Ming-Fan, Hsu, Chia-Lin, Huang, Chun-Ta, Wang, Jann-Yua, Hsieh, Shie-Lia, Yu, Chong-Je, Lee, Li-Na, Yang, Pan-Chyr
    BMC Infect Dis, 2013;13(0):45.
    Species: Human
    Sample Types: Serum
  8. DcR3 binds to ovarian cancer via heparan sulfate proteoglycans and modulates tumor cells response to platinum with corresponding alteration in the expression of BRCA1.
    BMC Cancer, 2012;12(0):176.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. The Decoy Receptor 3 (DcR3, TNFRSF6B) suppresses Th17 immune responses and is abundant in human cerebrospinal fluid.
    Authors: Mueller AM, Pedre X, Killian S, David M, Steinbrecher A
    J. Neuroimmunol., 2009;209(1):57-64.
    Species: Human
    Sample Types: Serum
  10. Comment on "Decoy receptor 3 expression in AsPC-1 human pancreatic adenocarcinoma cells via the phosphatidylinositol 3-kinase-, Akt-, and NF-kappa B-dependent pathway".
    Authors: Wang W, Sun W, Zhang L
    J. Immunol., 2009;183(4):2203; author.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Decoy receptor 3 levels in peripheral blood predict outcomes of acute respiratory distress syndrome.
    Authors: Chen CY, Yang KY, Chen MY, Chen HY, Lin MT, Lee YC, Perng RP, Hsieh SL, Yang PC, Chou TY
    Am. J. Respir. Crit. Care Med., 2009;180(8):751-60.
    Species: Human
    Sample Types: Plasma
  12. Decoy receptor 3 expression in AsPC-1 human pancreatic adenocarcinoma cells via the phosphatidylinositol 3-kinase-, Akt-, and NF-kappa B-dependent pathway.
    Authors: Chen PH, Yang CR
    J. Immunol., 2008;181(12):8441-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Epigenetic control of MHC class II expression in tumor-associated macrophages by decoy receptor 3.
    Authors: Chang YC, Chen TC, Lee CT, Yang CY, Wang HW, Wang CC, Hsieh SL
    Blood, 2008;111(10):5054-63.
    Species: Human
    Sample Types: Whole Cells

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