Porcine IL-6 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY686
Ancillary Products Available
Porcine IL-6 ELISA Standard Curve
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Product Details
Procedure
Citations (19)
FAQs
Supplemental Products
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Porcine IL-6 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
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)

Data Example

Porcine IL-6 ELISA Standard Curve

Product Datasheets

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

M-CSF, also known as CSF-1, is a four-alpha-helical-bundle cytokine that is the primary regulator of macrophage survival, proliferation and differentiation. M-CSF is also essential for the survival and proliferation of osteoclast progenitors. M-CSF also primes and enhances macrophage killing of tumor cells and microorganisms, regulates the release of cytokines and other inflammatory modulators from macrophages, and stimulates pinocytosis. M-CSF increases during pregnancy to support implantation and growth of the decidua and placenta. Sources of M-CSF include fibroblasts, activated macrophages, endometrial secretory epithelium, bone marrow stromal cells and activated endothelial cells. The M-CSF receptor (c-fms) transduces its pleotropic effects and mediates its endocytosis. M-CSF mRNAs of various sizes occur. Full length human M-CSF transcripts encode a 522 amino acid (aa) type I transmembrane (TM) protein with a 464 aa extracellular region, a 21 aa TM domain, and a 37 aa cytoplasmic tail that forms a 140 kDa covalent dimer. Differential processing produces two proteolytically cleaved, secreted dimers. One is an N- and O- glycosylated 86 kDa dimer, while the other is modified by both glycosylation and chondroitin-sulfate proteoglycan (PG) to generate a 200 kDa subunit. Although PG-modified M-CSF can circulate, it may be immobilized by attachment to type V collagen. Shorter transcripts encode M-CSF that lack cleavage and PG sites and produce an N-glycosylated 68 kDa TM dimer and a slowly produced 44 kDa secreted dimer. Although forms may vary in activity and half-life, all contain the N-terminal 150 aa portion that is necessary and sufficient for interaction with the M-CSF receptor. The first 223 aa of mature human M-CSF shares 88%, 86%, 81% and 74% aa identity with corresponding regions of dog, cow, mouse and rat M-CSF, respectively. Human M-CSF is active in the mouse, but mouse M-CSF is reported to be species-specific.

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 stimulatory factor-2; B-cell differentiation factor; BSF2; BSF-2; BSF2CTL differentiation factor; CDF; HGFHSFIFNB2Hybridoma growth factor; IFNB2; IFN-beta-2; IL6; IL-6; IL-6B-cell stimulatory factor 2; 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.

19 Citations: Showing 1 - 10
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  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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: Tissue Homogenates
  8. 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
  9. 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;8(12):e83182.
    Species: Porcine
    Sample Types: Plasma
  10. 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;8(4):e61904.
    Species: Porcine
    Sample Types: Plasma
  11. 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
  12. 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
  13. 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
  14. 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;37(3):1031-e4.
    Species: Porcine
    Sample Types: Plasma
  15. 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;138(8):1437-44.
    Species: Porcine
    Sample Types: Tissue Homogenates
  16. 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
  17. In vitro induction of mucosa-type dendritic cells by all-trans retinoic acid.
    Authors: Saurer L, McCullough KC, Summerfield A
    J. Immunol., 2007;179(6):3504-14.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  18. 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;14(10):836-44.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  19. 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;39(1):73-9.
    Species: Porcine
    Sample Types: Plasma

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