Human IL-21 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.
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
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: 0.1% BSA, 0.05% Tween 20 in Tris-buffered Saline (20mM Trizma base, 150mM NaCl) 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)
Product Datasheets
Preparation and Storage
Background: IL-21
IL-21 (Interleukin-21) is member of the family of cytokines that utilize the common gamma chain as a receptor subunit. This subunit is also a component of the receptors for IL-2, IL-4, IL-7, IL-9, and IL-15. IL-21 is produced by activated T follicular helper cells (Tfh), Th17 cells, and NKT cells. It exerts its biological effects through a heterodimeric receptor complex of the common gamma chain and the IL-21-specific IL-21 R. IL-21 supports Tfh cell development, immunoglobulin affinity maturation, plasma cell differentiation, B cell memory responses, and the migration of dendritic cells to draining lymph nodes. It costimulates the activation, proliferation, and survival of CD8+ T cells and NKT cells and promotes Th17 cell polarization. It blocks the generation of regulatory T cells and their suppressive effects on CD4+ T cells. In addition, IL-21 suppresses cutaneous hypersensitivity reactions by limiting allergen-specific IgE production and mast cell degranulation.
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-21 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.
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Citations: Showing 1 - 10
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Ex vivo T cell differentiation in adoptive immunotherapy manufacturing: Critical process parameters and analytical technologies
Authors: Chen, S;Nguyen, TD;Lee, KZ;Liu, D;
Biotechnology advances
Species: Human
Sample Types: Cell Culture Supernates
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Altered gene expression of miR-155 in peripheral blood mononuclear cells of Multiple sclerosis patients: Correlation with TH17 frequency, inflammatory cytokine profile and autoimmunity
Authors: Alipour, S;Amanallahi, P;Baradaran, B;Aghebati-Maleki, A;Soltani-Zangbar, MS;Aghebati-Maleki, L;
Multiple sclerosis and related disorders
Species: Human
Sample Types: Serum
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Interleukin-21 modulates balance between regulatory T cells and T-helper 17 cells in chronic hepatitis B virus infection
Authors: Cai, Y;Ji, H;Zhou, X;Zhao, K;Zhang, X;Pan, L;Shi, R;
BMC infectious diseases
Species: Human
Sample Types: Serum
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PD-1highCXCR5-CD4+ peripheral helper T�cells promote CXCR3+ plasmablasts in human acute viral infection
Authors: H Asashima, S Mohanty, M Comi, WE Ruff, KB Hoehn, P Wong, J Klein, C Lucas, I Cohen, S Coffey, N Lele, L Greta, K Raddassi, O Chaudhary, A Unterman, B Emu, SH Kleinstein, RR Montgomery, A Iwasaki, CS Dela Cruz, N Kaminski, AC Shaw, DA Hafler, TS Sumida
Cell Reports, 2023-01-02;0(0):111895.
Species: Human
Sample Types: Cell Culture Supernates
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Functional and structural characteristics of HLA-B*13:01-mediated specific T cells reaction in dapsone-induced drug hypersensitivity
Authors: H Jiang, CW Wang, Z Wang, Y Dai, Y Zhu, YS Lee, Y Cao, WH Chung, S Ouyang, H Wang
Journal of Biomedical Science, 2022-08-13;29(1):58.
Species: Human
Sample Types: Cell Culture Supernates
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Transient and durable T cell reactivity after COVID-19
Authors: A Martner, H Grauers Wi, A Törnell, J Ringlander, M Arabpour, M Lindh, M Lagging, S Nilsson, K Hellstrand
Oncogene, 2022-07-12;119(30):e2203659119.
Species: Human
Sample Types: Plasma
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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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Circulating Cytokines in Metastatic Breast Cancer Patients Select Different Prognostic Groups and Patients Who Might Benefit from Treatment beyond Progression
Authors: M Paccagnell, A Abbona, A Michelotti, E Geuna, F Ruatta, E Landucci, N Denaro, P Vanella, C Lo Nigro, D Galizia, M Merlano, O Garrone
Vaccines, 2022-01-05;10(1):.
Species: Human
Sample Types: Plasma
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Plasma interleukin-21 levels and genetic variants are associated with susceptibility to rheumatoid arthritis
Authors: Y Hao, L Xie, J Xia, Z Liu, B Yang, M Zhang
Bmc Musculoskeletal Disorders, 2021-03-05;22(1):246.
Species: Human
Sample Types: Plasma
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Defects of CTLA-4 Are Associated with Regulatory T Cells in Myasthenia Gravis Implicated by Intravenous Immunoglobulin Therapy
Authors: W Xu, M Ren, S Ghosh, K Qian, Z Luo, A Zhang, C Zhang, J Cui
Mediators Inflamm., 2020-02-14;2020(0):3645157.
Species: Human
Sample Types: Serum
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