Mouse IL-21 DuoSet ELISA

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Mouse IL-21 ELISA Standard Curve
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Product Details
Citations (31)
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Mouse IL-21 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sample Volume Required
100 µL
Assay Range
62.5 - 4,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
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 mouse IL-21. 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.

Normal Goat Serum: (Catalog # DY005)


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)

Normal Goat Serum: (Catalog # DY005)


Scientific Data

Mouse IL-21 ELISA Standard Curve

Product Datasheets

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Preparation and Storage

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: 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.

Long Name:
Interleukin 21
Entrez Gene IDs:
59067 (Human); 60505 (Mouse); 365769 (Rat); 442935 (Canine); 100344172 (Rabbit)
Alternate Names:
CVID11; IL21; IL-21; IL-21Za11interleukin-21; interleukin 21; interleukin-21 isoform; Za11

Assay Procedure


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 with NGS, 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 Mouse 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.

31 Citations: Showing 1 - 10
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  1. EBV/HHV-6A dUTPases contribute to myalgic encephalomyelitis/chronic fatigue syndrome pathophysiology by enhancing TFH cell differentiation and extrafollicular activities
    Authors: BS Cox, K Alharshawi, I Mena-Palom, WP Lafuse, ME Ariza
    JCI Insight, 2022-06-08;7(11):.
    Species: Mouse
    Sample Types: Tissue Homogenates
  2. Blimp-1 moulds the epigenetic architecture of IL-21-mediated autoimmune diseases through an autoregulatory circuit
    Authors: YW Liu, SH Fu, MW Chien, CY Hsu, MH Lin, JL Dong, RJ Lu, YJ Lee, PY Chen, CH Wang, HK Sytwu
    JCI Insight, 2022-06-08;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  3. ADP-ribosylating adjuvant reveals plasticity in cDC1 cells that drive mucosal Th17 cell development and protection against influenza virus infection
    Authors: M Arabpour, C Lebrero-Fe, K Schön, A Strömberg, V Börjesson, K Lahl, M Ballegeer, X Saelens, D Angeletti, W Agace, N Lycke
    Mucosal Immunology, 2022-04-13;0(0):.
    Species: Mouse
    Sample Types: Cell culture supernate
  4. Niclosamide suppresses the expansion of follicular helper T cells and alleviates disease severity in two murine models of lupus via STAT3
    Authors: SG Jang, J Lee, SM Hong, YS Song, MJ Kim, SK Kwok, ML Cho, SH Park
    Journal of Translational Medicine, 2021-02-25;19(1):86.
    Species: Mouse
    Sample Types: Serum
  5. Inhibition of Rho-Kinase Downregulates Th17 Cells and Ameliorates Hepatic Fibrosis by Schistosoma japonicum Infection
    Authors: W Zhou, Y Yang, C Mei, P Dong, S Mu, H Wu, Y Zhou, Y Zheng, F Guo, JQ Yang
    Cells, 2019-10-16;8(10):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  6. Ablation of RhoA impairs Th17 cell differentiation and alleviates house dust mite-triggered allergic airway inflammation
    Authors: JQ Yang, KW Kalim, Y Li, Y Zheng, F Guo
    J. Leukoc. Biol., 2019-07-01;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  7. IL-21/type I interferon interplay regulates neutrophil-dependent innate immune responses to Staphylococcus aureus
    Authors: R Spolski, EE West, P Li, S Veenbergen, S Yung, M Kazemian, J Oh, ZX Yu, AF Freeman, SM Holland, PM Murphy, WJ Leonard
    Elife, 2019-04-16;8(0):.
    Species: Mouse
    Sample Types: BALF
  8. Increases in helminth-induced IL-21 protein levels disappear upon sample freezing
    Authors: A Ariyaratne, EK Szabo, J Bowron, CAM Finney
    Cytokine, 2018-04-21;108(0):179-181.
    Species: Mouse
    Sample Types: Tissue Homogenates
  9. miR-146a modulates autoreactive Th17 cell differentiation and regulates organ-specific autoimmunity
    Authors: B Li, X Wang, IY Choi, YC Wang, S Liu, AT Pham, H Moon, DJ Smith, DS Rao, MP Boldin, L Yang
    J. Clin. Invest., 2017-09-05;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  10. Chitosan nanoparticle-based delivery of fused NKG2D-IL-21 gene suppresses colon cancer growth in mice
    Authors: L Tan, S Han, S Ding, W Xiao, Y Ding, L Qian, C Wang, W Gong
    Int J Nanomedicine, 2017-04-13;12(0):3095-3107.
    Species: Mouse
    Sample Types: Serum
  11. Bone marrow produces sufficient alloreactive natural killer (NK) cells in vivo to cure mice from subcutaneously and intravascularly injected 4T1 breast cancer
    Authors: Michel van Gelder
    Breast Cancer Res. Treat, 2016-12-03;0(0):.
    Species: Mouse
    Sample Types: Plasma
  12. Immune signatures of protective spleen memory CD8 T cells
    Sci Rep, 2016-11-24;6(0):37651.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  13. ER? in CD4+ T Cells Is Crucial for Ligand-Mediated Suppression of Central Nervous System Autoimmunity
    J Immunol, 2016-05-13;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  14. The adaptor TRAF5 limits the differentiation of inflammatory CD4(+) T cells by antagonizing signaling via the receptor for IL-6.
    Authors: Nagashima, Hiroyuki, Okuyama, Yuko, Asao, Atsuko, Kawabe, Takeshi, Yamaki, Satoshi, Nakano, Hiroyasu, Croft, Michael, Ishii, Naoto, So, Takanori
    Nat Immunol, 2014-03-30;15(5):449-56.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  15. Unique macrophages different from M1/M2 macrophages inhibit T cell mitogenesis while upregulating Th17 polarization.
    Authors: Tatano Y, Shimizu T, Tomioka H
    Sci Rep, 2014-02-20;4(0):4146.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  16. The activating protein 1 transcription factor basic leucine zipper transcription factor, ATF-like (BATF), regulates lymphocyte- and mast cell-driven immune responses in the setting of allergic asthma.
    Authors: Ubel C, Sopel N, Graser A, Hildner K, Reinhardt C, Zimmermann T, Rieker R, Maier A, Neurath M, Murphy K, Finotto S
    J Allergy Clin Immunol, 2013-11-28;133(1):198-206.e1-9.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  17. Retinoic acid prevents mesenteric lymph node dendritic cells from inducing IL-13-producing inflammatory Th2 cells.
    Authors: Yokota-Nakatsuma A, Takeuchi H, Ohoka Y, Kato C, Song S, Hoshino T, Yagita H, Ohteki T, Iwata M
    Mucosal Immunol, 2013-11-13;7(4):786-801.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  18. CXCR5(+) T helper cells mediate protective immunity against tuberculosis.
    Authors: Slight S, Rangel-Moreno J, Gopal R, Lin Y, Fallert Junecko B, Mehra S, Selman M, Becerril-Villanueva E, Baquera-Heredia J, Pavon L, Kaushal D, Reinhart T, Randall T, Khader S
    J Clin Invest, 2013-01-02;123(2):712-26.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  19. The encephalitogenicity of T(H)17 cells is dependent on IL-1- and IL-23-induced production of the cytokine GM-CSF.
    Authors: El-Behi M, Ciric B, Dai H
    Nat. Immunol., 2011-04-24;12(6):568-75.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  20. IL-21 Promotes Skin Recruitment of CD4+ Cells and Drives IFN-{gamma}-Dependent Epidermal Hyperplasia.
    Authors: Sarra M, Caruso R, Cupi ML, Monteleone I, Stolfi C, Campione E, Diluvio L, Mazzotta A, Botti E, Chimenti S, Costanzo A, MacDonald TT, Pallone F, Monteleone G
    J. Immunol., 2011-03-25;186(9):5435-42.
    Species: Mouse
    Sample Types: Tissue Homogenates
  21. Efficient induction of CCR9 on T cells requires coactivation of retinoic acid receptors and retinoid X receptors (RXRs): exaggerated T Cell homing to the intestine by RXR activation with organotins.
    Authors: Takeuchi H, Yokota A, Ohoka Y, Kagechika H, Kato C, Song SY, Iwata M
    J. Immunol., 2010-09-29;185(9):5289-99.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  22. Critical role of the disintegrin metalloprotease ADAM17 for intestinal inflammation and regeneration in mice.
    Authors: Chalaris A, Adam N, Sina C, Rosenstiel P, Lehmann-Koch J, Schirmacher P, Hartmann D, Cichy J, Gavrilova O, Schreiber S, Jostock T, Matthews V, Hasler R, Becker C, Neurath MF, Reiss K, Saftig P, Scheller J, Rose-John S
    J. Exp. Med., 2010-07-05;207(8):1617-24.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  23. The AP-1 transcription factor Batf controls T(H)17 differentiation.
    Authors: Schraml BU, Hildner K, Ise W, Lee WL, Smith WA, Solomon B, Sahota G, Sim J, Mukasa R, Cemerski S, Hatton RD, Stormo GD, Weaver CT, Russell JH, Murphy TL, Murphy KM
    Nature, 2009-07-05;460(7253):405-9.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  24. The induction of antibody production by IL-6 is indirectly mediated by IL-21 produced by CD4+ T cells.
    Authors: Dienz O, Eaton SM, Bond JP, Neveu W, Moquin D, Noubade R, Briso EM, Charland C, Leonard WJ, Ciliberto G, Teuscher C, Haynes L, Rincon M
    J. Exp. Med., 2009-01-12;206(1):69-78.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  25. Local IL-21 promotes the therapeutic activity of effector T cells by decreasing regulatory T cells within the tumor microenvironment.
    Authors: Kim-Schulze S, Kim HS, Fan Q, Kim DW, Kaufman HL
    Mol. Ther., 2008-11-25;17(2):380-8.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  26. A fundamental role for interleukin-21 in the generation of T follicular helper cells.
    Authors: Vogelzang A, McGuire HM, Yu D, Sprent J, Mackay CR, King C
    Immunity, 2008-07-03;29(1):127-37.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  27. Regulation of humoral and cellular gut immunity by lamina propria dendritic cells expressing Toll-like receptor 5.
    Authors: Uematsu S, Fujimoto K, Jang MH, Yang BG, Jung YJ, Nishiyama M, Sato S, Tsujimura T, Yamamoto M, Yokota Y, Kiyono H, Miyasaka M, Ishii KJ, Akira S
    Nat. Immunol., 2008-05-30;9(7):769-76.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  28. IL-21 is produced by Th17 cells and drives IL-17 production in a STAT3-dependent manner.
    Authors: Wei L, Laurence A, Elias KM, O'Shea JJ
    J. Biol. Chem., 2007-09-20;282(48):34605-10.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  29. Interleukin-21 activates cytotoxic T lymphocytes and natural killer cells to generate antitumor response in mouse renal cell carcinoma.
    Authors: Kumano M, Hara I, Furukawa J, Oniki S, Nagai H, Miyake H, Fujisawa M
    J. Urol., 2007-08-16;178(4):1504-9.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  30. IL-21 is produced by NKT cells and modulates NKT cell activation and cytokine production.
    Authors: Coquet JM, Kyparissoudis K, Pellicci DG, Besra G, Berzins SP, Smyth MJ, Godfrey DI
    J. Immunol., 2007-03-01;178(5):2827-34.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  31. Interleukin-2 gene variation impairs regulatory T cell function and causes autoimmunity.
    Authors: Yamanouchi J, Rainbow D, Serra P, Howlett S, Hunter K, Garner VE, Gonzalez-Munoz A, Clark J, Veijola R, Cubbon R, Chen SL, Rosa R, Cumiskey AM, Serreze DV, Gregory S, Rogers J, Lyons PA, Healy B, Smink LJ, Todd JA, Peterson LB, Wicker LS, Santamaria P
    Nat. Genet., 2007-02-04;39(3):329-37.
    Species: Mouse
    Sample Types: Cell Culture Supernates


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Mouse IL-21 DuoSet ELISA
By William Jackson on 11/13/2019
Sample Tested: Serum