Mouse sTNF RII/TNFRSF1B DuoSet ELISA Summary
* 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 mouse sTNF RII/TNFRSF1B. 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
Product Datasheets
Preparation and Storage
Background: TNF RII/TNFRSF1B
TNF RII (Tumor Necrosis Factor Receptor II), also known as TNFRSF1B, p75/p80, and CD120b, is a widely expressed receptor for membrane-associated TNF-alpha and Lymphotoxin-alpha. Its activation initiates pro-inflammatory and pro-survival responses via NFkB-dependent signaling pathways, although it may also induce apoptosis.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- 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.
- 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.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- 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 Mouse sTNF RII/TNFRSF1B 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.
8
Citations: Showing 1 - 8
Filter your results:
Filter by:
-
Acute Kidney Injury-Induced Circulating TNFR1/2 Elevations Correlate with Persistent Kidney Injury and Progression to Fibrosis
Authors: Arthanarisami, A;Komaru, Y;Katsouridi, C;Schumacher, J;Verges, DK;Ning, L;Abdelmageed, MM;Herrlich, A;Kefaloyianni, E;
Cells
Species: Mouse
Sample Types: Serum, Urine
-
Differential organ-specific inflammatory response to progranulin in high-fat diet-fed mice
Authors: M Murakoshi, T Gohda, E Adachi, S Ichikawa, S Hagiwara, Y Suzuki
Scientific Reports, 2021-01-13;11(1):1194.
Species: Mouse
Sample Types: Urine
-
Mesenchymal stem cells inhibit cutaneous radiation-induced fibrosis by suppressing chronic inflammation.
Authors: Horton J, Hudak K, Chung E, White A, Scroggins B, Burkeen J, Citrin D
Stem Cells, 2013-10-01;31(10):2231-41.
Species: Mouse
Sample Types: Tissue Homogenates
-
Tumor necrosis factor-alpha from macrophages enhances LPS-induced clara cell expression of keratinocyte-derived chemokine.
Authors: Elizur A, Adair-Kirk TL, Kelley DG, Griffin GL, deMello DE, Senior RM
Am. J. Respir. Cell Mol. Biol., 2007-08-02;38(1):8-15.
Species: Mouse
Sample Types: Cell Culture Supernates
-
Interleukin 10 regulates inflammatory cytokine synthesis to protect against lipopolysaccharide-induced abortion and fetal growth restriction in mice.
Authors: Robertson SA, Care AS, Skinner RJ
Biol. Reprod., 2007-01-10;76(5):738-48.
Species: Mouse
Sample Types: Serum
-
Essential role for IL-10 in resistance to lipopolysaccharide-induced preterm labor in mice.
Authors: Robertson SA, Skinner RJ, Care AS
J. Immunol., 2006-10-01;177(7):4888-96.
Species: Mouse
Sample Types: Serum
-
Anti-tumor necrosis factor alpha F(ab')2 antibody fragments protect in murine polymicrobial sepsis: concentration and early intervention are fundamental to the outcome.
Authors: Marquez-Velasco R, Bojalil R, Buelna A, Flores-Guzman F, Estevez-Ramirez J, Laguna J, Hernandez AM, Diaz-Quinonez A, Paniagua-Solis JF
Inflamm. Res., 2006-09-01;55(9):378-84.
Species: Mouse
Sample Types: Serum
-
Increased pulmonary responses to acute ozone exposure in obese db/db mice.
Authors: Lu FL, Johnston RA, Flynt L, Theman TA, Terry RD, Schwartzman IN, Lee A, Shore SA
Am. J. Physiol. Lung Cell Mol. Physiol., 2005-12-22;290(5):L856-65.
Species: Mouse
Sample Types: Serum
FAQs
No product specific FAQs exist for this product, however you may
View all ELISA FAQsReviews for Mouse sTNF RII/TNFRSF1B DuoSet ELISA
There are currently no reviews for this product. Be the first to review Mouse sTNF RII/TNFRSF1B DuoSet ELISA and earn rewards!
Have you used Mouse sTNF RII/TNFRSF1B DuoSet ELISA?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥2500 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image
