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Porcine TNF-alpha DuoSet ELISA

Catalog #: DY690B
38 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY690B
Ancillary Products Available
DY008B: DuoSet ELISA Ancillary Reagent Kit 2
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
Porcine TNF-alpha ELISA Standard Curve
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Citations (38)
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Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Porcine TNF-alpha DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
31.2 - 2,000 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 TNF-alpha. 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

N/A Porcine TNF-alpha ELISA Standard Curve View Larger

Porcine TNF-alpha 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: TNF-alpha

Tumor necrosis factor alpha (TNF-α), also known as cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, apoptosis, and immune system development. TNF-α is produced by a wide variety of immune and epithelial cell types. Human TNF-α consists of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD). Within the ECD, human TNF-α shares 97% aa sequence identity with rhesus and 71% - 92% with bovine, canine, cotton rat, equine, feline, mouse, porcine, and rat TNF-α. The 26 kDa type 2 transmembrane protein is assembled intracellularly to form a noncovalently linked homotrimer. Ligation of this complex induces reverse signaling that promotes lymphocyte costimulation but diminishes monocyte responsiveness.

Cleavage of membrane bound TNF-α by TACE/ADAM17 releases a 55 kDa soluble trimeric form of TNF-α. TNF-α trimers bind the ubiquitous TNF RI and the hematopoietic cell-restricted TNF RII, both of which are also expressed as homotrimers. TNF-α regulates lymphoid tissue development through control of apoptosis. It also promotes inflammatory responses by inducing the activation of vascular endothelial cells and macrophages. TNF-α is a key cytokine in the development of several inflammatory disorders. It contributes to the development of type 2 diabetes through its effects on insulin resistance and fatty acid metabolism.

Long Name:
Tumor Necrosis Factor alpha
Entrez Gene IDs:
7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 102139631 (Cynomolgus Monkey); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)
Alternate Names:
APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; TNFA; TNF-A; TNFalpha; TNF-alpha; TNF-alphacachectin; TNFATNF, macrophage-derived; TNFG1F; TNFSF1A; TNFSF2; TNFSF2TNF superfamily, member 2; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor; tumor necrosis factor-alpha

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 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 TNF-alpha 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.

38 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. Citrulline supplementation exacerbates sepsis severity in infected preterm piglets via early induced immunosuppression
    Authors: Zhong, J;Johansen, SH;Bæk, O;Nguyen, DN;
    The Journal of nutritional biochemistry
    Species: Porcine
    Sample Types: Plasma
  3. Comparative analysis of the interactions of different Streptococcus suis strains with monocytes, granulocytes and the complement system in porcine blood
    Authors: Zhu, H;Müller, U;Baums, CG;Öhlmann, S;
    Veterinary research
    Species: Porcine
    Sample Types: Plasma
  4. Dietary seaweed (Saccharina latissima) supplementation in pigs induces localized immunomodulatory effects and minor gut microbiota changes during intestinal helminth infection
    Authors: Bonde, CS;Mejer, H;Myhill, LJ;Zhu, L;Jensen, P;Büdeyri Gökgöz, N;Krych, L;Nielsen, DS;Skovgaard, K;Thamsborg, SM;Williams, AR;
    Scientific reports
    Species: Porcine
    Sample Types: Cell Culture Supernates
  5. 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
  6. Comparative effects of soy protein concentrate, enzyme-treated soybean meal, and fermented soybean meal replacing animal protein supplements in feeds on growth performance and intestinal health of nursery pigs
    Authors: Deng, Z;Duarte, ME;Kim, SY;Hwang, Y;Kim, SW;
    Journal of animal science and biotechnology
    Species: Porcine
    Sample Types: Tissue Homogenates
  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. Infant Formula Based on Milk Fat Affects Immune Development in Both Normal Birthweight and Fetal Growth Restricted Neonatal Piglets
    Authors: O Bæk, K Skadborg, T Muk, C Amdi, PMH Heegaard, T Thymann, DN Nguyen
    Nutrients, 2021-09-22;13(10):.
    Species: Porcine
    Sample Types: Plasma
  10. PIIKA 2.5: Enhanced quality control of peptide microarrays for kinome analysis
    Authors: C Denomy, C Lazarou, D Hogan, A Facciuolo, E Scruten, A Kusalik, S Napper
    PLoS ONE, 2021-09-10;16(9):e0257232.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  11. Early Immune Initiation by Porcine Cells following Toxoplasma gondii Infection versus TLR Ligation
    Authors: B Hamid, J Schlosser-, L Bechtold, F Ebner, S Rausch, S Hartmann
    Microorganisms, 2021-08-28;9(9):.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  12. 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
  13. 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
  14. Proline-rich antimicrobial peptide Api137 is bactericidal in porcine blood infected ex vivo with a porcine or human Klebsiella pneumoniae strain
    Authors: AK Krieger, D Knappe, S Öhlmann, L Mayer, IB Eder, G Köller, R Hoffmann, K Rieckmann, CG Baums
    Journal of global antimicrobial resistance, 2020-12-26;24(0):127-135.
    Species: Porcine
    Sample Types: Plasma
  15. 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
  16. 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
  17. 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
  18. Substitution of warthog NF-kappaB motifs into RELA of domestic pigs is not sufficient to confer resilience to African swine fever virus
    Authors: S McCleary, R Strong, RR McCarthy, JC Edwards, EL Howes, LM Stevens, PJ Sánchez-Co, A Núñez, S Watson, AJ Mileham, SG Lillico, C Tait-Burka, C Proudfoot, M Ballantyne, CBA Whitelaw, F Steinbach, HR Crooke
    Sci Rep, 2020-06-02;10(1):8951.
    Species: Porcine
    Sample Types: Serum
  19. 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
  20. Beneficial Effect of Mildly Pasteurized Whey Protein on Intestinal Integrity and Innate Defense in Preterm and Near-Term Piglets
    Authors: M Navis, V Muncan, PT Sangild, L Møller Wil, PJ Koelink, ME Wildenberg, E Abrahamse, T Thymann, RM van Elburg, IB Renes
    Nutrients, 2020-04-17;12(4):.
    Species: Porcine
    Sample Types: Tissue Homogenates
  21. Effect of polymorphisms in porcine guanylate-binding proteins on host resistance to PRRSV infection in experimentally challenged pigs
    Authors: A Khatun, S Nazki, CG Jeong, S Gu, SUS Mattoo, SI Lee, MS Yang, B Lim, KS Kim, B Kim, KT Lee, CK Park, SM Lee, WI Kim
    Vet. Res., 2020-02-19;51(1):14.
    Species: Porcine
    Sample Types: Serum
  22. 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
  23. Addition of dairy lipids and probiotic Lactobacillus fermentum in infant formula programs gut microbiota and entero-insular axis in adult minipigs
    Authors: M Lemaire, S Dou, A Cahu, M Formal, L Le Normand, V Romé, I Nogret, S Ferret-Ber, M Rhimi, I Cuinet, C Canlet, M Tremblay-F, P Le Ruyet, C Baudry, P Gérard, I Le Huërou-, S Blat
    Sci Rep, 2018-08-03;8(1):11656.
    Species: Human
    Sample Types: Cell Cultue Supernates
  24. Plant synthetic GP4 and GP5 proteins from porcine reproductive and respiratory syndrome virus elicit immune responses in pigs
    Authors: CH An, S Nazki, SC Park, YJ Jeong, JH Lee, SJ Park, A Khatun, WI Kim, YI Park, JC Jeong, CY Kim
    Planta, 2018-01-08;0(0):.
    Species: Porcine
    Sample Types: Serum
  25. 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
  26. 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
  27. Acute social stress-induced immunomodulation in pigs high and low responders to ACTH
    Authors: Blandine Lieubeau
    Physiol. Behav., 2016-11-17;169(0):1-8.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  28. 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
  29. Antimicrobial and immunomodulatory activities of PR-39 derived peptides.
    Authors: Veldhuizen E, Schneider V, Agustiandari H, van Dijk A, Tjeerdsma-van Bokhoven J, Bikker F, Haagsman H
    PLoS ONE, 2014-04-22;9(4):e95939.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  30. The peripheral blood transcriptome reflects variations in immunity traits in swine: towards the identification of biomarkers.
    Authors: Mach N, Gao Y, Lemonnier G, Lecardonnel J, Oswald I, Estelle J, Rogel-Gaillard C
    BMC Genomics, 2013-12-17;14(0):894.
    Species: Porcine
    Sample Types: Whole Blood
  31. 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
  32. CD27 expression discriminates porcine T helper cells with functionally distinct properties.
    Authors: Reutner K, Leitner J, Mullebner A, Ladinig A, Essler S, Duvigneau J, Ritzmann M, Steinberger P, Saalmuller A, Gerner W
    Vet Res, 2013-03-11;44(0):18.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  33. 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., 2012-04-24;43(1):35.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  34. 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
  35. 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
  36. 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
  37. 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
  38. Inflammation increases vulnerability to hypoxia in newborn piglets: effect of reoxygenation with 21% and 100% O2.
    Authors: Lyng K, Braakhuis M, Froen JF, Stray-Pedersen B, Saugstad OD
    Am. J. Obstet. Gynecol., 2005-04-01;192(4):1172-8.
    Species: Porcine
    Sample Types: Plasma

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