Human IL-7 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 human IL-7. 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: IL-7
IL-7 (Interleukin-7) is a cytokine that plays important roles in lymphocyte differentiation, proliferation, and survival. IL-7 is produced by stromal epithelial cells of the thymus, bone marrow, and intestines. It signals through a receptor complex composed of IL-7 R alpha/CD127 and the Common gamma Chain. gamma c is also a subunit of the receptors for IL-2, -4, -9, -15, and -21. IL-7 contributes to the maintenance of all naïve and memory T cells. It is required for optimal T cell-dendritic cell interaction. In mouse, IL-7 activation of IL-7 R alpha is critical for both T cell and B cell lineage development, while in humans, it is required for T cell but not for B cell development.
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 Human IL-7 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.
11
Citations: Showing 1 - 10
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Ventilatory support and inflammatory peptides in hospitalised patients with COVID-19: A prospective cohort trial
Authors: Gysan, MR;Milacek, C;Bal, C;Zech, A;Brugger, J;Milos, RI;Antoniewicz, L;Idzko, M;Gompelmann, D;
PloS one
Species: Human
Sample Types: Serum
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Cytokine Adjuvants IL-7 and IL-15 Improve Humoral Responses of a SHIV LentiDNA Vaccine in Animal Models
Authors: LA Leroy, A Mac Donald, A Kandlur, D Bose, P Xiao, J Gagnon, F Villinger, Y Chebloune
Vaccines, 2022-03-17;10(3):.
Species: Human
Sample Types: Cell Culture Supernates
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Characterization of memory T cell subsets and common gamma-chain cytokines in convalescent COVID-19 individuals
Authors: A Rajamanick, N Pavan Kuma, AN Pandiaraj, N Selvaraj, S Munisankar, RM Renji, V Venkataram, M Murhekar, JWV Thangaraj, SK Muthusamy, GK Chethrapil, T Bhatnagar, M Ponnaiah, S Ramasamy, S Velusamy, S Babu
Journal of leukocyte biology, 2022-03-08;0(0):.
Species: Human
Sample Types: Plasma
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WIN 55,212-2 shows anti-inflammatory and survival properties in human iPSC-derived cardiomyocytes infected with SARS-CoV-2
Authors: LGHS Aragão, JT Oliveira, JR Temerozo, MA Mendes, JA Salerno, CSG Pedrosa, T Puig-Pijua, CP Veríssimo, IM Ornelas, T Torquato, G Vitória, CQ Sacramento, N Fintelman-, S da Silva G, V Cardoso So, LRQ Souza, K Karmirian, L Goto-Silva, D Biagi, EM Cruvinel, R Dariolli, DR Furtado, PT Bozza, HL Borges, TML Souza, MZP Guimarães, SK Rehen
PeerJ, 2021-10-08;9(0):e12262.
Species: Human
Sample Types: Cell Culture Supernates
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Neutrophils promote T-cell activation through the regulated release of CD44-bound Galectin-9 from the cell surface during HIV infection
Authors: G Dunsmore, EP Rosero, S Shahbaz, DM Santer, J Jovel, P Lacy, S Houston, S Elahi
PloS Biology, 2021-08-19;19(8):e3001387.
Species: Human
Sample Types: Plasma
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Altered plasma levels of &betaC and &gammaC chain cytokines and post-treatment modulation in tuberculous lymphadenitis
Authors: GR Kathamuthu, K Moideen, R Sridhar, D Baskaran, S Babu
Cytokine, 2020-12-17;138(0):155405.
Species: Human
Sample Types: Plasma
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The tuberculosis vaccine candidate Bacillus Calmette-Guerin DeltaureC::hly coexpressing human interleukin-7 or -18 enhances antigen-specific T cell responses in mice.
Authors: Rao M, Vogelzang A, Kaiser P, Schuerer S, Kaufmann S, Gengenbacher M
PLoS ONE, 2013-11-13;8(11):e78966.
Species: Bacteria
Sample Types: Cell Culture Supernates
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Effects of in utero arsenic exposure on child immunity and morbidity in rural Bangladesh.
Authors: Raqib R, Ahmed S, Sultana R, Wagatsuma Y, Mondal D, Hoque AM, Nermell B, Yunus M, Roy S, Persson LA, Arifeen SE, Moore S, Vahter M
Toxicol. Lett., 2009-01-09;185(3):197-202.
Species: Human
Sample Types: Milk
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Human bronchial intraepithelial T cells produce interferon-gamma and stimulate epithelial cells.
Authors: Hirosako S, Goto E, Fujii K, Tsumori K, Hirata N, Tsumura S, Kamohara H, Kohrogi H
Clin. Exp. Immunol., 2008-11-26;155(2):266-74.
Species: Human
Sample Types: Cell Culture Supernates
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Islet transplantation in patients with autoimmune diabetes induces homeostatic cytokines that expand autoreactive memory T cells.
Authors: Monti P, Scirpoli M, Maffi P, Ghidoli N, De Taddeo F, Bertuzzi F, Piemonti L, Falcone M, Secchi A, Bonifacio E
J. Clin. Invest., 2008-05-01;118(5):1806-14.
Species: Human
Sample Types: Serum
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Human T cells constitutively express IL-15 that promotes ex vivo T cell homeostatic proliferation through autocrine/juxtacrine loops.
Authors: Miranda-Carus ME, Benito-Miguel M, Llamas MA, Balsa A, Martin-Mola E
J. Immunol., 2005-09-15;175(6):3656-62.
Species: Human
Sample Types: Cell Culture Supernates
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293 T cells were transfected with human IL-7 expressing plasmids and IL-7 in the cell supernatants were analyzed by ELISA.
