Home / IL-6 / Porcine IL-6 DuoSet ELISA

Porcine IL-6 DuoSet ELISA

Catalog #: DY686
36 Citations 2 Reviews Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY686
Ancillary Products Available
DY008B: DuoSet ELISA Ancillary Reagent Kit 2
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
Porcine IL-6 ELISA Standard Curve
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Product Details
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Citations (36)
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Reviews (2)
Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Porcine IL-6 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
125.0 - 8,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
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-6. 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

N/A Porcine IL-6 ELISA Standard Curve View Larger

Porcine IL-6 ELISA Standard Curve

Product Datasheets

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Product Datasheet
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Preparation and Storage

Shipping
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-6

Interleukin 6 (IL-6) is a pleiotropic, alpha -helical, 22 - 28 kDa phosphorylated and variably glycosylated cytokine that plays important roles in the acute phase reaction, inflammation, hematopoiesis, bone metabolism, and cancer progression (1-5). Mature porcine IL-6 is 183 amino acids (aa) in length and shares 58%, 39% and 39% aa sequence identity with human, mouse, and rat IL-6, respectively (6). Cells known to express IL-6 include CD8+ T cells, fibroblasts, synoviocytes, adipocytes, osteoblasts, megakaryocytes, endothelial cells (under the influence of endothelins), sympathetic neurons, cerebral cortex neurons, adrenal medulla chromaffin cells, retinal pigment cells, mast cells, keratinocytes, Langerhans cells, fetal and adult astrocytes, neutrophils, monocytes, eosinophils, colonic epithelial cells, B1 B cells and pancreatic islet beta cells (2, 7-29). IL-6 production is generally correlated with cell activation and is normally kept in control by glucocorticoids, catecholamines, and secondary sex steroids (2). Normal human circulating IL-6 is in the 1 pg/mL range, with slight elevations during the menstrual cycle, modest elevations in certain cancers, and large elevations after surgery (30-34). 

IL-6 induces signaling through a cell surface heterodimeric receptor complex composed of a ligand binding subunit (IL-6 Ra) and a signal transducing subunit (gp130). IL-6 binds to IL-6 R alpha, triggering IL-6 R alpha association with gp130, and gp130 dimerization (35). gp130 is also a component of the receptors for CLC, CNTF, CT-1, IL-11, IL-27, LIF, and OSM (36). Soluble forms of IL-6 R alpha are generated by both alternative splicing and proteolytic cleavage (5). In a mechanism known as trans-signaling, complexes of soluble IL-6 and IL-6 R alpha elicit responses from gp130- expressing cells that lack cell surface IL-6 R alpha (5). Trans-signaling enables a wider range of cell types to respond to IL-6, as the expression of gp130 is ubiquitous, while that of IL-6 R alpha is predominantly restricted to hepatocytes, monocytes, and resting lymphocytes (2, 5). Soluble splice forms of gp130 block trans-signaling from IL-6/IL-6 R alpha but not from other cytokines that use gp130 as a co-receptor (5, 37). 
IL-6, along with TNF-alpha and IL-1, drives the acute inflammatory response. IL-6 is almost solely responsible for fever and the acute phase response in the liver, and it is important in the transition from acute inflammation to either acquired immunity or chronic inflammatory disease (1-5). When dysregulated, it contributes to chronic inflammation in conditions such as obesity, insulin resistance, inflammatory bowel disease, arthritis, and sepsis (2, 5). IL-6 modulates bone resorption and is a major effector of inflammatory joint destruction in rheumatoid arthritis through its promotion of Th17 cell development and activity (1). It contributes to atherosclerotic plaque development and destabilization as well as the development of inflammation-associated carcinogenesis (1, 2). IL-6 can also function as an anti-inflammatory molecule, as in skeletal muscle where it is secreted in response to exercise (2). In addition, it enhances hematopoietic stem cell proliferation and the differentiation of memory B cells and plasma cells (38).

Long Name:
Interleukin 6
Entrez Gene IDs:
3569 (Human); 16193 (Mouse); 24498 (Rat); 399500 (Porcine); 280826 (Bovine); 403985 (Canine); 102138971 (Cynomolgus Monkey); 100034196 (Equine); 493687 (Feline); 463288 (Primate); 100008733 (Rabbit)
Alternate Names:
B-cell differentiation factor; B-cell stimulatory factor 2; BSF2; BSF-2; CDF; CTL differentiation factor ; HSF; hybridoma growth factor; IFNB2; IFN-beta-2; IL6; IL-6; Interferon beta-2; interleukin 6 (interferon, beta 2); interleukin BSF-2; interleukin-6; MGI-2A

Assay Procedure

GENERAL ELISA PROTOCOL

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, 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-6 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.

36 Citations: Showing 1 - 10
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  1. The lipopolysaccharide structure affects the detoxifying ability of intestinal alkaline phosphatases
    Authors: Vermeire, B;Walsh, M;Cox, E;Devriendt, B;
    BMC veterinary research
    Species: Porcine
    Sample Types: Cell Culture Supernates
  2. Predicting cytokine kinetics during sepsis; a modelling framework from a porcine sepsis model with live Escherichia coli
    Authors: Bahnasawy, SM;Skorup, P;Hanslin, K;Lipcsey, M;Friberg, LE;Nielsen, EI;
    Cytokine
    Species: Porcine
    Sample Types: Plasma
  3. Efficacy of Feed Additive Containing Bentonite and Enzymatically Hydrolyzed Yeast on Intestinal Health and Growth of Newly Weaned Pigs under Chronic Dietary Challenges of Fumonisin and Aflatoxin
    Authors: Deng, Z;Jang, KB;Jalukar, S;Du, X;Kim, SW;
    Toxins
    Species: Porcine
    Sample Types: Tissue Homogenate
  4. Anti-inflammatory properties of antiangiogenic fucoidan in retinal pigment epithelium cells
    Authors: P Dörschmann, C Seeba, T Thalenhors, J Roider, A Klettner
    Heliyon, 2023-04-10;9(4):e15202.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  5. Anti-inflammatory properties of antiangiogenic fucoidan in retinal pigment epithelium cells
    Authors: P Dörschmann, C Seeba, T Thalenhors, J Roider, A Klettner
    Heliyon, 2023;9(4):e15202.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  6. Pulmonary inflammatory response and immunomodulation to multiple trauma and hemorrhagic shock in pigs
    Authors: MA Oestreich, K Seidel, W Bertrams, HH Müller, M Sassen, T Steinfeldt, H Wulf, B Schmeck
    PLoS ONE, 2022-12-07;17(12):e0278766.
    Species: Porcine
    Sample Types: BALF
  7. Use of a novel "Split" ventilation system in bench and porcine modeling of acute respiratory distress syndrome
    Authors: P Geoghegan, J Clarke, G Hogan, A Keogh, H Marsh, K Donnelly, N McEvoy, A Doolan, SF Madden, I Martin-Loe, M Power, JG Laffey, GF Curley
    Physiological Reports, 2022-09-01;10(17):e15452.
    Species: Porcine
    Sample Types: BALF
  8. Glucose supply and glycolysis inhibition shape the clinical fate of Staphylococcus epidermidis-infected preterm newborns
    Authors: T Muk, A Brunse, NL Henriksen, K Aasmul-Ols, DN Nguyen
    JCI Insight, 2022-06-08;0(0):.
    Species: Porcine
    Sample Types: Plasma
  9. Human-relevant near-organ neuromodulation of the immune system via the splenic nerve
    Authors: M Donegà, CT Fjordbakk, J Kirk, DM Sokal, I Gupta, GE Hunsberger, A Crawford, S Cook, J Viscasilla, TR Stathopoul, JA Miranda, WJ Dopson, D Goodwin, A Rowles, P McGill, A McSloy, D Werling, J Witheringt, DJ Chew, JD Perkins
    Proceedings of the National Academy of Sciences of the United States of America, 2021-05-18;118(20):.
    Species: Porcine
    Sample Types: Plasma
  10. Lung-protective ventilation increases cerebral metabolism and non-inflammatory brain injury in porcine experimental sepsis
    Authors: A Nyberg, E Gremo, J Blixt, J Sperber, A Larsson, M Lipcsey, A Pikwer, M Castegren
    Bmc Neuroscience, 2021-04-29;22(1):31.
    Species: Porcine
    Sample Types: Plasma
  11. A comprehensive assessment of multi-system responses to a renal inoculation of uropathogenic E. coli in swine
    Authors: MH Tiba, BM McCracken, RP Dickson, JA Nemzek, CI Colmenero, DC Leander, TL Flott, RC Daniels, KE Konopka, JS VanEpps, KA Stringer, KR Ward
    PLoS ONE, 2020-12-11;15(12):e0243577.
    Species: Porcine
    Sample Types: Whole Blood
  12. Pre-exposure to mechanical ventilation and endotoxemia increases Pseudomonas aeruginosa growth in lung tissue during experimental porcine pneumonia
    Authors: J Sperber, A Nyberg, A Krifors, P Skorup, M Lipcsey, M Castegren
    PLoS ONE, 2020-10-27;15(10):e0240753.
    Species: Porcine
    Sample Types: BALF
  13. The antisecretory peptide AF-16 may modulate tissue edema but not inflammation in experimental peritonitis induced sepsis
    Authors: A Barrueta T, J van der He, I Blokhin, F Massaro, HA Hansson, R Feinstein, A Larsson, A Larsson, J Tenhunen
    PLoS ONE, 2020-08-21;15(8):e0232302.
    Species: Porcine
    Sample Types: Plasma
  14. Activation of the NLRP3 Inflammasome by Particles from the Echinococcus granulosus Laminated Layer
    Authors: C Casaravill, Á Pittini, D Rückerl, JE Allen, Á Díaz
    Infect. Immun., 2020-08-19;88(9):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  15. Lung-protective ventilation suppresses systemic and hepatic vein levels of cell-free DNA in porcine experimental post-operative sepsis
    Authors: A Nyberg, A Larsson, J Jylhävä, M Hurme, J Sperber, M Lipcsey, M Castegren
    BMC Pulm Med, 2020-07-31;20(1):206.
    Species: Porcine
    Sample Types: Plasma
  16. 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), 2020-05-27;12(6):.
    Species: Porcine
    Sample Types: Whole Tissue
  17. Analysis of Porcine Pro- and Anti-Inflammatory Cytokine Induction by S. suis In Vivo and In Vitro
    Authors: FS Hohnstein, M Meurer, N de Buhr, M von Köckri, CG Baums, G Alber, N Schütze
    Pathogens, 2020-01-03;9(1):.
    Species: Porcine
    Sample Types: Whole Cells
  18. 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., 2019-11-08;50(1):91.
    Species: Porcine
    Sample Types: BALF
  19. 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., 2019-01-29;230(0):101-109.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  20. 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, 2018-06-09;15(1):180.
    Species: Porcine
    Sample Types: Tissue Homogenates
  21. 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, 2017-12-19;7(1):17861.
    Species: Porcine
    Sample Types: Tissue Homogenates
  22. 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., 2017-08-29;13(1):274.
    Species: Porcine
    Sample Types: BALF
  23. 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, 2017-06-12;7(1):3224.
    Species: Porcine
    Sample Types: Tissue Culture Supernates
  24. 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, 2017-02-28;13(1):64.
    Species: Porcine
    Sample Types: BALF
  25. 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, 2014-09-25;45(0):95.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  26. Lung protective ventilation induces immunotolerance and nitric oxide metabolites in porcine experimental postoperative sepsis.
    Authors: Sperber J, Lipcsey M, Larsson A, Larsson A, Sjolin J, Castegren M
    PLoS ONE, 2013-12-12;8(12):e83182.
    Species: Porcine
    Sample Types: Plasma
  27. Immunopathogenesis of severe acute respiratory disease in Zaire ebolavirus-infected pigs.
    Authors: Nfon, Charles, Leung, Anders, Smith, Greg, Embury-Hyatt, Carissa, Kobinger, Gary, Weingartl, Hana M
    PLoS ONE, 2013-04-23;8(4):e61904.
    Species: Porcine
    Sample Types: Plasma
  28. 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., 2012-04-19;43(1):30.
    Species: Porcine
    Sample Types: Serum
  29. 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., 2012-02-07;32(5):221-9.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  30. 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., 2010-03-10;17(5):728-34.
    Species: Porcine
    Sample Types: Tissue Homogenates
  31. Inflammatory and circulatory effects of the reduction of endotoxin concentration in established porcine endotoxemic shock--a model of endotoxin elimination.
    Authors: Carlsson M, Lipcsey M, Larsson A, Tano E, Rubertsson S, Eriksson M, Sjolin J
    Crit. Care Med., 2009-03-01;37(3):1031-e4.
    Species: Porcine
    Sample Types: Plasma
  32. Early administration of probiotics alters bacterial colonization and limits diet-induced gut dysfunction and severity of necrotizing enterocolitis in preterm pigs.
    Authors: Siggers RH, Siggers J, Boye M, Thymann T, Molbak L, Leser T, Jensen BB, Sangild PT
    J. Nutr., 2008-08-01;138(8):1437-44.
    Species: Porcine
    Sample Types: Tissue Homogenates
  33. 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., 2008-07-21;27(9):1016-22.
    Species: Porcine
    Sample Types: BALF
  34. In vitro induction of mucosa-type dendritic cells by all-trans retinoic acid.
    Authors: Saurer L, McCullough KC, Summerfield A
    J. Immunol., 2007-09-15;179(6):3504-14.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  35. Toll-like receptor 7 and MyD88 knockdown by lentivirus-mediated RNA interference to porcine dendritic cell subsets.
    Authors: Alves MP, Neuhaus V, Guzylack-Piriou L, Ruggli N, McCullough KC, Summerfield A
    Gene Ther., 2007-03-01;14(10):836-44.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  36. Neutrophil and small intestinal lymphocyte migration after Salmonella typhimurium infection: impact of fermentable fiber.
    Authors: Milo LA, Correa-Matos NJ, Donovan SM, Tappenden KA
    J. Pediatr. Gastroenterol. Nutr., 2004-07-01;39(1):73-9.
    Species: Porcine
    Sample Types: Plasma

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Reviews for Porcine IL-6 DuoSet ELISA

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Porcine IL-6 DuoSet ELISA
By Anonymous on 03/10/2022
Sample Tested: Porcine Cell Culture Supernates

Works ok. As a reagent diluent RPMI medium can be used to dilute standard curve and samples.

Porcine IL-6 DuoSet ELISA DY686
Porcine IL-6 DuoSet ELISA
By Anonymous on 08/10/2020
Sample Tested: Cell culture supernatant,Porcine Cell Culture Supernates

easy and reliable assay

Porcine IL-6 DuoSet ELISA DY686