Human IL-15 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-15. 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-15
IL-15 (Interleukin 15) is a cytokine that induces or enhances the differentiation, maintenance, or activation of multiple T cell subsets including NK, NKT, Th17, Treg, and CD8+ memory cells. It also induces dendritic cell differentiation and inflammatory activation, exhibits anti-tumor activity, and inhibits the deposition of lipid in adipocytes. Complexes of IL-15 with cell surface IL-15 R alpha interact with complexes of IL-2 R beta and the Common gamma Chain on adjacent cells. This transpresentation mechanism enables cells to respond to IL-15 even if they do not express IL-15 R alpha. Ligation of membrane-associated IL-15/IL-15 R alpha complexes also induces reverse signaling that promotes activation of the IL-15/IL-15 R alpha expressing cells. The activity of circulating IL-15 is limited by its association with soluble IL-15 R alpha.
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-15 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.
19
Citations: Showing 1 - 10
Filter your results:
Filter by:
-
Mild Cognitive Impairment Is Associated with Enhanced Activation of Th17 Lymphocytes in Non-Alcoholic Fatty Liver Disease
Authors: Fiorillo, A;Gallego, JJ;Casanova-Ferrer, F;Giménez-Garzó, C;Urios, A;Ballester, MP;Durbán, L;Rios, MP;Megías, J;San Miguel, T;Kosenko, E;Escudero-García, D;Benlloch, S;Felipo, V;Montoliu, C;
International journal of molecular sciences
Species: Human
Sample Types: Plasma
-
Secreted Fas decoys enhance the antitumor activity of engineered and bystander T cells in Fas ligand-expressing solid tumors
Authors: P Bajgain, AG Torres Cha, K Balasubram, L Fleckenste, P Lulla, HE Heslop, J Vera, AM Leen
Oncogene, 2022-11-02;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
-
IL-15/IL-15Ralpha in SJS/TEN: Relevant Expression of IL15 and IL15RA in Affected Skin
Authors: T Bellón, O González-V, E Sendagorta, V Lerma, J Martínez D, C Martínez, G Servera, C González-H, L Cachafeiro, JA Lorente, R Cabañas, P Herranz, F de Abajo, PIELenRed
Biomedicines, 2022-08-02;10(8):.
Species: Human
Sample Types: Blister Fluid
-
HydrAd: A Helper-Dependent Adenovirus Targeting Multiple Immune Pathways for Cancer Immunotherapy
Authors: A Rosewell S, C Porter, G Biegert, L Jatta, M Suzuki
Oncogene, 2022-06-02;14(11):.
Species: Human
Sample Types: Cell Culture Supernates
-
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
-
Opposing roles for sMAdCAM and IL-15 in COVID-19 associated cellular immune pathology
Authors: AK Singh, N Kasarpalka, S Bhowmick, G Paradkar, M Talreja, K Shah, A Tiwari, H Palav, S Kaginkar, R Kulkarni, A Patil, V Kalsurkar, S Agrawal, J Shastri, R Dere, R Bharmal, SD Mahale, VM Bhor, V Patel
Journal of leukocyte biology, 2022-01-24;0(0):.
Species: Human
Sample Types: Plasma
-
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
-
DeSUMOylase SENP7-Mediated Epithelial Signaling Triggers Intestinal Inflammation via Expansion of Gamma-Delta T Cells
Authors: A Suhail, ZA Rizvi, P Mujagond, SA Ali, P Gaur, M Singh, V Ahuja, A Awasthi, CV Srikanth
Cell Rep, 2019-12-10;29(11):3522-3538.e7.
Species: Human
Sample Types: Cell Culture Supernates
-
Rheumatoid arthritis bone marrow environment supports Th17 response.
Authors: Ewa Kuca-Warna, Weronika Kurowska, Monika Prochorec, Anna Radzikows, Tomasz Burakowsk, Urszula Skalska, Magdalena Massalska, Magdalena Pleba?czy, Barbara Ma?dyk-No, Iwona S?owi?ska, Robert Gasik, W?odzimierz Ma?li?ski
Arthritis Research & Therapy, 2017-12-08;0(0):1478-6362.
Species: Human
Sample Types: Plasma
-
Priming of Human Resting NK Cells by Autologous M1 Macrophages via the Engagement of IL-1beta, IFN-beta, and IL-15 Pathways.
Authors: Mattiola I, Pesant M, Tentorio P, Molgora M, Marcenaro E, Lugli E, Locati M, Mavilio D
J Immunol, 2015-08-14;195(6):2818-28.
Species: Human
Sample Types: Cell Culture Supernates
-
IL15RA drives antagonistic mechanisms of cancer development and immune control in lymphocyte-enriched triple-negative breast cancers.
Authors: Marra P, Mathew S, Grigoriadis A, Wu Y, Kyle-Cezar F, Watkins J, Rashid M, De Rinaldis E, Hessey S, Gazinska P, Hayday A, Tutt A
Cancer Res, 2014-06-30;74(17):4908-21.
Species: Human
Sample Types: Cell Lysates
-
Interleukin-15 contributes to the regulation of murine adipose tissue and human adipocytes.
Authors: Barra NG, Reid S, MacKenzie R
Obesity (Silver Spring), 2009-12-17;18(8):1601-7.
Species: Human
Sample Types: Complex Sample Type
-
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
-
How a cytokine is chaperoned through the secretory pathway by complexing with its own receptor: lessons from interleukin-15 (IL-15)/IL-15 receptor alpha.
Authors: Duitman EH, Orinska Z, Bulanova E, Paus R, Bulfone-Paus S
Mol. Cell. Biol., 2008-05-27;28(15):4851-61.
Species: Human
Sample Types: Cell Culture Supernates
-
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
-
Soluble Interleukin IL-15Ralpha is generated by alternative splicing or proteolytic cleavage and forms functional complexes with IL-15.
Authors: Bulanova E, Budagian V, Duitman E, Orinska Z, Krause H, Ruckert R, Reiling N, Bulfone-Paus S
J. Biol. Chem., 2007-02-27;282(18):13167-79.
Species: Human
Sample Types: Cell Culture Supernates
-
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
-
IL-15 and the initiation of cell contact-dependent synovial fibroblast-T lymphocyte cross-talk in rheumatoid arthritis: effect of methotrexate.
Authors: Miranda-Carus ME, Balsa A, Benito-Miguel M, Perez de Ayala C, Martin-Mola E
J. Immunol., 2004-07-15;173(2):1463-76.
Species: Human
Sample Types: Cell Culture Supernates
-
Stimulation of T cell autoreactivity by anomalous expression of NKG2D and its MIC ligands in rheumatoid arthritis.
Authors: Groh V, Bruhl A, El-Gabalawy H, Nelson JL, Spies T
Proc. Natl. Acad. Sci. U.S.A., 2003-07-23;100(16):9452-7.
Species: Human
Sample Types: Serum
FAQs
No product specific FAQs exist for this product, however you may
View all ELISA FAQsReviews for Human IL-15 DuoSet ELISA
Average Rating: 5 (Based on 2 Reviews)
Have you used Human IL-15 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
Filter by:
Detection of IL-15 secreted by human T cells engineered to express IL-15 (PMC617), compared to normal T cells.
