Porcine IL-1 beta/IL-1F2 DuoSet ELISA

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Porcine IL-1 beta / IL-1F2 ELISA Standard Curve
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Citations (16)
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Porcine IL-1 beta/IL-1F2 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 porcine IL-1ß/IL-1F2. 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

Block Buffer: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Reagent Diluent: 0.1% BSA, 0.05% Tween 20 in Tris-buffered Saline (20 mM Trizma base, 150 mM NaCI) 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

Porcine IL-1 beta / IL-1F2 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-1 beta/IL-1F2

IL-1 beta (Interleukin-1 beta) is produced in response to inflammatory agents, infections, or microbial endotoxins. It plays a central role in immune and inflammatory responses, bone remodeling, fever, carbohydrate metabolism, and GH/IGF-I physiology. IL-1 beta dysregulation is implicated in many pathological conditions including sepsis, rheumatoid arthritis, inflammatory bowel disease, acute and chronic myelogenous leukemia, insulin-dependent diabetes mellitus, atherosclerosis, neuronal injury, and aging-related diseases. Intracellular cleavage of the IL-1 beta precursor by Caspase-1/ICE is a key step in the inflammatory response. IL-1 beta binds to IL-1 RI and IL-1 RII. The IL-1 receptor accessory protein (IL-1 RAcP) associates with IL-1 RI and is required for IL-1 RI signal transduction, and IL-1ra is a secreted molecule that functions as a competitive inhibitor of IL-1 bioactivity.

Long Name:
Interleukin 1 beta
Entrez Gene IDs:
3553 (Human); 16176 (Mouse); 24494 (Rat); 397122 (Porcine); 403974 (Canine); 100034237 (Equine); 100135556 (Guinea Pig)
Alternate Names:
catabolin; IL1 beta; IL-1 beta; IL-1; IL1B; IL-1b; IL1-BETA; IL-1F2; IL1F2IL-1 beta; interleukin 1, beta; interleukin-1 beta; preinterleukin 1 beta; pro-interleukin-1-beta

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 of Block Buffer 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, 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 Porcine IL-1 beta/IL-1F2 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.

16 Citations: Showing 1 - 10
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  1. The Effect of Zearalenone on the Cytokine Environment, Oxidoreductive Balance and Metabolism in Porcine Ileal Peyer's Patches
    Authors: K Obremski, W Trybowski, P Wojtacha, M Gaj?cka, J Tyburski, ? Zielonka
    Toxins (Basel), 2020;12(6):.
    Species: Porcine
    Sample Types: Whole Tissue
  2. Efficacy of three innovative bacterin vaccines against experimental infection with Mycoplasma hyopneumoniae
    Authors: AMF Matthijs, G Auray, F Boyen, A Schoos, A Michiels, O García-Nic, GT Barut, C Barnier-Qu, V Jakob, N Collin, B Devriendt, A Summerfiel, F Haesebrouc, D Maes
    Vet. Res., 2019;50(1):91.
    Species: Porcine
    Sample Types: BALF
  3. Classical swine fever virus non-structural proteins modulate Toll-like receptor signaling pathways in porcine monocyte-derived macrophages
    Authors: Z Cao, Q Yang, M Zheng, H Lv, K Kang, Y Zhang
    Vet. Microbiol., 2019;230(0):101-109.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  4. Necrotizing enterocolitis is associated with acute brain responses in preterm pigs
    Authors: J Sun, X Pan, LI Christians, XL Yuan, K Skovgaard, DEW Chatterton, SS Kaalund, F Gao, PT Sangild, S Pankratova
    J Neuroinflammation, 2018;15(1):180.
    Species: Porcine
    Sample Types: Tissue Homogenates
  5. Host-pathogen interplay at primary infection sites in pigs challenged with Actinobacillus pleuropneumoniae
    Authors: EL Sassu, J Frömbling, JC Duvigneau, I Miller, A Müllebner, AM Gutiérrez, T Grunert, M Patzl, A Saalmüller, A von Altroc, A Menzel, M Ganter, J Spergser, M Hewicker-T, J Verspohl, M Ehling-Sch, I Hennig-Pau
    BMC Vet. Res, 2017;13(1):64.
    Species: Porcine
    Sample Types: BALF
  6. Cardiac Depression in Pigs after Multiple Trauma - Characterization of Posttraumatic Structural and Functional Alterations
    Authors: M Kalbitz, S Schwarz, B Weber, B Bosch, J Pressmar, FM Hoenes, CK Braun, K Horst, TP Simon, R Pfeifer, P Störmann, H Hummler, F Gebhard, HC Pape, M Huber-Lang, F Hildebrand, TREAT Rese
    Sci Rep, 2017;7(1):17861.
    Species: Porcine
    Sample Types: Whole Tissue
  7. Efficacy of one dose vaccination against experimental infection with two Mycoplasma hyopneumoniae strains
    Authors: A Michiels, I Arsenakis, F Boyen, R Krejci, F Haesebrouc, D Maes
    BMC Vet. Res., 2017;13(1):274.
    Species: Porcine
    Sample Types: BALF
  8. Early Gut Microbiota Intervention Suppresses DSS-Induced Inflammatory Responses by Deactivating TLR/NLR Signalling in Pigs
    Authors: Y Xiao, H Yan, H Diao, B Yu, J He, J Yu, P Zheng, X Mao, Y Luo, D Chen
    Sci Rep, 2017;7(1):3224.
    Species: Porcine
    Sample Types: Tissue Culture Supernates
  9. Human milk oligosaccharide effects on intestinal function and inflammation after preterm birth in pigs
    Authors: Stine B Bering
    J. Nutr. Biochem., 2016;40(0):141-154.
    Species: Porcine
    Sample Types: Tissue Homogenates
  10. The immune response against Chlamydia suis genital tract infection partially protects against re-infection.
    Authors: De Clercq E, Devriendt B, Yin L, Chiers K, Cox E, Vanrompay D
    Vet Res, 2014;45(0):95.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  11. N(pro) of Classical Swine Fever Virus Prevents Type I Interferon-Mediated Priming of Conventional Dendritic Cells for Enhanced Interferon-alpha Response.
    Authors: Husser L, Ruggli N, Summerfield A
    J. Interferon Cytokine Res., 2012;32(5):221-9.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  12. Characterization of homologous and heterologous adaptive immune responses in porcine reproductive and respiratory syndrome virus infection.
    Authors: Diaz I, Gimeno M, Darwich L, Navarro N, Kuzemtseva L, Lopez S, Galindo I, Segales J, Martin M, Pujols J, Mateu E
    Vet. Res., 2012;43(1):30.
    Species: Porcine
    Sample Types: Serum
  13. The fungal T-2 toxin alters the activation of primary macrophages induced by TLR-agonists resulting in a decrease of the inflammatory response in the pig.
    Vet. Res., 2012;43(1):35.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  14. Cytokine protein expression levels in tracheobronchial lymph node homogenates of pigs infected with pseudorabies virus.
    Authors: Miller LC, Zanella EL, Waters WR
    Clin. Vaccine Immunol., 2010;17(5):728-34.
    Species: Porcine
    Sample Types: Tissue Homogenates
  15. Impact of topical cooling solution and prediction of pulmonary graft viability from non-heart-beating donors.
    Authors: Inci I, Arni S, Inci D, Zhai W, Hillinger S, Leskosek B, Vogt P, Weder W
    J. Heart Lung Transplant., 2008;27(9):1016-22.
    Species: Porcine
    Sample Types: BALF
  16. Expression of porcine endometrial prostaglandin synthase during the estrous cycle and early pregnancy, and following endocrine disruption of pregnancy.
    Authors: Ashworth MD, Ross JW, Hu J, White FJ, Stein DR, Desilva U, Johnson GA, Spencer TE, Geisert RD
    Biol. Reprod., 2006;74(6):1007-15.
    Species: Porcine
    Sample Types: Uterine Lavage Fluid


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Porcine IL-1 beta/IL-1F2 DuoSet ELISA
By Anonymous on 01/21/2020
Sample Tested: porcine macrophages

The standard curve works well