Porcine TNF-alpha Quantikine ELISA Kit

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PTA00
TNF-alpha ELISA Kit ELISA
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Citations (45)
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Porcine TNF-alpha Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
Sensitivity
5 pg/mL
Assay Range
23.4 - 1,500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant porcine TNF-alpha
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Porcine TNF-alpha Immunoassay is a 4.5 hour solid phase ELISA designed to measure porcine TNF-alpha in cell culture supernates, serum, and plasma samples. It contains E. coli-expressed recombinant porcine TNF-alpha and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately measure recombinant porcine TNF-alpha. Results obtained using natural porcine TNF-alpha show dose response curves that are parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural porcine TNF-alpha.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 79 260 646 88 279 679
Standard Deviation 4.9 9.2 27 8.8 24 44
CV% 6.2 3.5 4.2 10 8.6 6.5

Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 104 358 880 106 348 866
Standard Deviation 7.2 13 36 9.7 32 72
CV% 6.9 3.6 4.1 9.2 9.2 8.3

Recovery

The recovery of porcine TNF-alpha spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=6) 99 91-105
EDTA Plasma (n=6) 98 83-111
Heparin Plasma (n=4) 91 82-99
Serum (n=6) 105 91-120

Linearity

To assess the linearity of the assay, six samples containing and/or spiked with various concentrations of porcine TNF-alpha in each matrix were diluted with the appropriate Calibrator Diluent and then assayed.
Porcine TNF-alpha ELISA Linearity

Data Examples

Porcine TNF-alpha ELISA Cell Culture Supernate Standard Curve

Porcine TNF-alpha ELISA Serum/Plasma Standard Curve

Product Datasheets

Preparation and Storage

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); 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; 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
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Porcine TNF-alpha Quantikine ELISA Kit

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.

45 Citations: Showing 1 - 10
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  1. Modulation of the somatotropic axis, adiponectin and cytokine secretion during highly pathogenic porcine reproductive and respiratory syndrome virus type 1 (HP-PRRSV-1) infection
    Authors: R Saleri, V Cavalli, L Ferrari, G Ogno, E Canelli, P Martelli, P Borghetti
    Res. Vet. Sci., 2019;124(0):263-269.
    Species: Porcine
    Sample Types: Plasma
  2. Inflammatory markers before and after farrowing in healthy sows and in sows affected with postpartum dysgalactia syndrome
    Authors: M Kaiser, M Jacobson, PH Andersen, P Bækbo, JJ Cerón, J Dahl, D Escribano, S Jacobsen
    BMC Vet. Res., 2018;14(1):83.
    Species: Porcine
    Sample Types: Serum
  3. Pulmonary Effects of Remote Ischemic Preconditioning in a Porcine Model of Ventilation-induced Lung Injury
    Authors: A Bergmann, T Schilling, G Hedenstier, K Ahlgren, A Larsson, M Kretzschma, A Kozian, T Hachenberg
    Respir Physiol Neurobiol, 2018;0(0):.
    Species: Porcine
    Sample Types: Tissue Homogenates
  4. Medium-Chain Triglycerides Attenuate Liver Injury in Lipopolysaccharide-Challenged Pigs by Inhibiting Necroptotic and Inflammatory Signaling Pathways
    Authors: L Zhang, X Wang, S Chen, S Wang, Z Tu, G Zhang, H Zhu, X Li, J Xiong, Y Liu
    Int J Mol Sci, 2018;19(11):.
    Species: Porcine
    Sample Types: Serum
  5. Blood pressure targeting by partial REBOA is possible in severe hemorrhagic shock in pigs and produces less circulatory, metabolic and inflammatory sequelae than total REBOA
    Authors: M Sadeghi, TM Hörer, D Forsman, EM Dogan, K Jansson, C Kindler, P Skoog, KF Nilsson
    Injury, 2018;0(0):.
    Species: Porcine
    Sample Types: Serum
  6. Glutamate alleviates intestinal injury, maintains mTOR and suppresses TLR4 and NOD signaling pathways in weanling pigs challenged with lipopolysaccharide
    Authors: Q Qin, X Xu, X Wang, H Wu, H Zhu, Y Hou, B Dai, X Liu, Y Liu
    Sci Rep, 2018;8(1):15124.
    Species: Porcine
    Sample Types: Plasma
  7. Neuroimmunological Implications of Subclinical Lipopolysaccharide from Salmonella Enteritidis
    Authors: A Miko?ajczy, D Z?otkowska
    Int J Mol Sci, 2018;19(10):.
    Species: Porcine
    Sample Types: Serum
  8. Butyrate modifies intestinal barrier function in IPEC-J2 cells through a selective upregulation of tight junction proteins and activation of the Akt signaling pathway
    Authors: H Yan, KM Ajuwon
    PLoS ONE, 2017;12(6):e0179586.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  9. Altered Cytokine Expression and Barrier Properties after In Vitro Infection of Porcine Epithelial Cells with Enterotoxigenic Escherichia coli and Probiotic Enterococcus faecium
    Authors: M Kern, D Günzel, JR Aschenbach, K Tedin, A Bondzio, U Lodemann
    Mediators Inflamm., 2017;2017(0):2748192.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  10. Inhibition of CD34+ cell migration by matrix metalloproteinase-2 during acute myocardial ischemia, counteracted by ischemic preconditioning
    Authors: D Lukovic, K Zlabinger, A Gugerell, A Spannbauer, N Pavo, L Mandic, DT Weidenauer, S Kastl, C Kaun, A Posa, I Sabdyushev, J Winkler, M Gyöngyösi
    F1000Res, 2016;5(0):2739.
    Species: Porcine
    Sample Types: Serum
  11. Septic porcine blood does not further activate coagulation during in vitro membrane oxygenation
    Authors: Rolf Rossaint
    Eur J Cardiothorac Surg, 2016;0(0):.
    Species: Porcine
    Sample Types: Plasma
  12. An Immunomodulatory Device Improves Insulin Resistance in Obese Porcine Model of Metabolic Syndrome
    Authors: Angela J Westover
    J Diabetes Res, 2016;2016(0):3486727.
    Species: Porcine
    Sample Types: Plasma
  13. Inhalation therapy with the synthetic TIP-like peptide AP318 attenuates pulmonary inflammation in a porcine sepsis model.
    Authors: Hartmann E, Ziebart A, Thomas R, Liu T, Schad A, Tews M, Moosmann B, Kamuf J, Duenges B, Thal S, David M
    BMC Pulm Med, 2015;15(0):7.
    Species: Porcine
    Sample Types: Plasma
  14. Sulforaphane epigenetically regulates innate immune responses of porcine monocyte-derived dendritic cells induced with lipopolysaccharide.
    Authors: Qu X, Proll M, Neuhoff C, Zhang R, Cinar M, Hossain M, Tesfaye D, Grosse-Brinkhaus C, Salilew-Wondim D, Tholen E, Looft C, Holker M, Schellander K, Uddin M
    PLoS ONE, 2015;10(3):e0121574.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  15. Low tidal volume pressure support versus controlled ventilation in early experimental sepsis in pigs.
    Authors: Ziebart A, Hartmann E, Thomas R, Liu T, Duenges B, Schad A, Bodenstein M, Thal S, David M
    Respir Res, 2014;15(0):101.
    Species: Porcine
    Sample Types: Plasma
  16. Efficacy of linezolid compared to vancomycin in an experimental model of pneumonia induced by methicillin-resistant Staphylococcus aureus in ventilated pigs.
    Authors: Martinez-Olondris P, Rigol M, Soy D, Guerrero L, Agusti C, Quera MA, Li Bassi G, Esperatti M, Luque N, Liapikou M, Filella X, Marco F, de la Bellacasa JP, Torres A
    Crit. Care Med., 2012;40(1):162-8.
    Species: Porcine
    Sample Types: Serum
  17. Risks Associated with High-Dose Lactobacillus rhamnosus in an Escherichia coli Model of Piglet Diarrhoea: Intestinal Microbiota and Immune Imbalances.
    Authors: Li XQ, Zhu YH, Zhang HF, Yue Y, Cai ZX, Lu QP, Zhang L, Weng XG, Zhang FJ, Zhou D, Yang JC, Wang JF
    PLoS ONE, 2012;7(7):e40666.
    Species: Porcine
    Sample Types: Serum
  18. Ornithodoros moubata complement inhibitor is an equally effective C5 inhibitor in pigs and humans.
    Authors: Barratt-Due A, Thorgersen EB, Lindstad JK, Pharo A, Lissina O, Lambris JD, Nunn MA, Mollnes TE
    J. Immunol., 2011;187(9):4913-9.
    Species: Porcine
    Sample Types: Plasma
  19. Plasma concentrations of inflammatory cytokines rise rapidly during ECMO-related SIRS due to the release of preformed stores in the intestine.
    Authors: McILwain RB, Timpa JG, Kurundkar AR
    Lab. Invest., 2010;90(1):128-39.
    Species: Porcine
    Sample Types: Plasma
  20. CD14 inhibition efficiently attenuates early inflammatory and hemostatic responses in Escherichia coli sepsis in pigs.
    Authors: Thorgersen EB, Hellerud BC, Nielsen EW, Barratt-Due A, Fure H, Lindstad JK, Pharo A, Fosse E, Tonnessen TI, Johansen HT, Castellheim A, Mollnes TE
    FASEB J., 2010;24(3):712-22.
    Species: Porcine
    Sample Types: Plasma
  21. Dissecting the effects of lipopolysaccharides from nonlipopolysaccharide molecules in experimental porcine meningococcal sepsis.
    Authors: Hellerud BC, Nielsen EW, Thorgersen EB, Lindstad JK, Pharo A, Tonnessen TI, Castellheim A, Mollnes TE, Brandtzaeg P
    Crit. Care Med., 2010;38(6):1467-74.
    Species: Porcine
    Sample Types: Plasma
  22. Autologous mesenchymal stem cells produce reverse remodelling in chronic ischaemic cardiomyopathy.
    Authors: Schuleri KH, Feigenbaum GS, Centola M, Weiss ES, Zimmet JM, Turney J, Kellner J, Zviman MM, Hatzistergos KE, Detrick B, Conte JV, McNiece I, Steenbergen C, Lardo AC, Hare JM
    Eur. Heart J., 2009;30(22):2722-32.
    Species: Porcine
    Sample Types: Serum
  23. Effect of linezolid compared with glycopeptides in methicillin-resistant Staphylococcus aureus severe pneumonia in piglets.
    Authors: Luna CM, Bruno DA, Garcia-Morato J, Mann KC, Risso Patron J, Sagardia J, Absi R, Garcia Bottino M, Marchetti D, Famiglietti A, Baleztena M, Biancolini C
    Chest, 2009;135(6):1564-71.
    Species: Porcine
    Sample Types: Serum
  24. Inhibition of complement and CD14 attenuates the Escherichia coli-induced inflammatory response in porcine whole blood.
    Authors: Thorgersen EB, Pharo A, Haverson K, Axelsen AK, Gaustad P, Kotwal GJ, Sfyroera G, Mollnes TE
    Infect. Immun., 2009;77(2):725-32.
    Species: Porcine
    Sample Types: Whole Blood
  25. Cardiac Function and the Proinflammatory Cytokine Response After Recovery From Cardiac Arrest in Swine.
    Authors: Niemann JT, Rosborough JP, Youngquist S, Shah AP, Lewis RJ, Phan QT, Filler SG
    J. Interferon Cytokine Res., 2009;0(0):.
    Species: Porcine
    Sample Types: Plasma
  26. Metabolic activity of the enteric microbiota influences the fatty acid composition of murine and porcine liver and adipose tissues.
    Authors: Wall R, Ross RP, Shanahan F, O'Mahony L, O'Mahony C, Coakley M, Hart O, Lawlor P, Quigley EM, Kiely B, Fitzgerald GF, Stanton C
    Am. J. Clin. Nutr., 2009;89(5):1393-401.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  27. Blood concentrations of the cytokines IL-1beta, IL-6, IL-10, TNF-alpha and IFN-gamma during experimentally induced swine dysentery.
    Authors: Kruse R, Essen-Gustavsson B, Fossum C, Jensen-Waern M
    Acta Vet. Scand., 2008;50(0):32.
    Species: Porcine
    Sample Types: Serum
  28. Inducible nitric oxide synthase expression and cardiomyocyte dysfunction during sustained moderate ischemia in pigs.
    Authors: Heinzel FR, Gres P, Boengler K, Duschin A, Konietzka I, Rassaf T, Snedovskaya J, Meyer S, Skyschally A, Kelm M, Heusch G, Schulz R
    Circ. Res., 2008;103(10):1120-7.
    Species: Porcine
    Sample Types: Cell Lysates
  29. Protective effect of Sivelestat in a porcine hepatectomy model prepared using an intermittent Pringle method.
    Authors: Shimoda M, Iwasaki Y, Okada T, Sawada T, Kubota K
    Eur. J. Pharmacol., 2008;587(1):248-52.
    Species: Porcine
    Sample Types: Serum
  30. Cytokine secretion depends on Galalpha(1,3)Gal expression in a pig-to-human whole blood model.
    Authors: Saethre M, Schneider MK, Lambris JD, Magotti P, Haraldsen G, Seebach JD, Mollnes TE
    J. Immunol., 2008;180(9):6346-53.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  31. Effects of glucocorticoids in ventilated piglets with severe pneumonia.
    Authors: Sibila O, Luna CM, Agusti C, Baquero S, Gando S, Patron JR, Morato JG, Absi R, Bassi N, Torres A
    Eur. Respir. J., 2008;32(4):1037-46.
    Species: Porcine
    Sample Types: Serum
  32. Involvement of both tumor necrosis factor-alpha-induced necrosis and p53-mediated caspase-dependent apoptosis in nephrotoxicity of cisplatin.
    Authors: Yano T, Itoh Y, Matsuo M, Kawashiri T, Egashira N, Oishi R
    Apoptosis, 2007;12(10):1901-9.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  33. Experimental severe Pseudomonas aeruginosa pneumonia and antibiotic therapy in piglets receiving mechanical ventilation.
    Authors: Luna CM, Baquero S, Gando S, Patron JR, Morato JG, Sibila O, Absi R, Famiglietti A, Vay CA, Von Stecher F, Agusti C, Torres A
    Chest, 2007;132(2):523-31.
    Species: Porcine
    Sample Types: Serum
  34. Bidirectional role of tumor necrosis factor-alpha in coronary microembolization: progressive contractile dysfunction versus delayed protection against infarction.
    Authors: Skyschally A, Gres P, Hoffmann S, Haude M, Erbel R, Schulz R, Heusch G
    Circ. Res., 2007;100(1):140-6.
    Species: Porcine
    Sample Types: Tissue Homogenates
  35. Experimental Pseudomonas aeruginosa pneumonia: evaluation of the associated inflammatory response.
    Authors: Sibila O, Agusti C, Torres A, Baquero S, Gando S, Patron JR, Morato JG, Goffredo DH, Bassi N, Luna CM
    Eur. Respir. J., 2007;30(6):1167-72.
    Species: Porcine
    Sample Types: Serum
  36. The fibrin-derived peptide Bbeta15-42 is cardioprotective in a pig model of myocardial ischemia-reperfusion injury.
    Authors: Roesner JP, Petzelbauer P, Koch A, Mersmann J, Zacharowski PA, Boehm O, Reingruber S, Pasteiner W, Mascher D, Wolzt M, Barthuber C, Noldge-Schomburg GE, Scheeren TW, Zacharowski K
    Crit. Care Med., 2007;35(7):1730-5.
    Species: Porcine
    Sample Types: Plasma
  37. Activated protein C downregulates p38 mitogen-activated protein kinase and improves clinical parameters in an in-vivo model of septic shock.
    Authors: Nold MF, Nold-Petry CA, Fischer D, Richter B, Blaheta R, Pfeilschifter J, Muhl H, Schranz D, Veldman A
    Thromb. Haemost., 2007;98(5):1118-26.
    Species: Porcine
    Sample Types: Plasma
  38. Dietary fatty acid composition rather than vitamin E supplementation influence ex vivo cytokine and eicosanoid response of porcine alveolar macrophages.
    Authors: Moller S, Lauridsen C
    Cytokine, 2006;35(1):6-12.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  39. Proangiogenic effects of protease-activated receptor 2 are tumor necrosis factor-alpha and consecutively Tie2 dependent.
    Authors: Zhu T, Sennlaub F, Beauchamp MH, Fan L, Joyal JS, Checchin D, Nim S, Lachapelle P, Sirinyan M, Hou X, Bossolasco M, Rivard GE, Heveker N, Chemtob S
    Arterioscler. Thromb. Vasc. Biol., 2006;26(4):744-50.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  40. Role of oxidative stress in remodeling of the myocardial microcirculation in hypertension.
    Authors: Zhu XY, Daghini E, Chade AR, Rodriguez-Porcel M, Napoli C, Lerman A, Lerman LO
    Arterioscler. Thromb. Vasc. Biol., 2006;26(8):1746-52.
    Species: Porcine
    Sample Types: Plasma
  41. Conjugated linoleic acid attenuates the production and gene expression of proinflammatory cytokines in weaned pigs challenged with lipopolysaccharide.
    Authors: Changhua L, Jindong Y, Defa L, Lidan Z, Shiyan Q, Jianjun X
    J. Nutr., 2005;135(2):239-44.
    Species: Porcine
    Sample Types: Serum
  42. The effect of warm humidified CO2 on the dissipation of residual gas following laparoscopy in piglets.
    Authors: Glew PA, Campher MJ, Pearson K, Schofield JC, Davey AK
    J Am Assoc Gynecol Laparosc, 2004;11(2):204-10.
    Species: Porcine
    Sample Types: Serum
  43. Deciphering the involvement of innate immune factors in the development of the host response to PRRSV vaccination.
    Authors: Royaee AR, Husmann RJ, Dawson HD, Calzada-Nova G, Schnitzlein WM, Zuckermann FA, Lunney JK
    Vet. Immunol. Immunopathol., 2004;102(3):199-216.
    Species: Porcine
    Sample Types: Whole Cells
  44. Extracellular ubiquitin inhibits the TNF-alpha response to endotoxin in peripheral blood mononuclear cells and regulates endotoxin hyporesponsiveness in critical illness.
    Authors: Majetschak M, Krehmeier U, Bardenheuer M, Denz C, Quintel M, Voggenreiter G, Obertacke U
    Blood, 2003;101(5):1882-90.
    Species: Porcine
    Sample Types: Plasma
  45. Ex vivo adenoviral vector gene delivery results in decreased vector-associated inflammation pre- and post-lung transplantation in the pig.
    Authors: Yeung JC, Wagnetz D, Cypel M, Rubacha M, Koike T, Chun YM, Hu J, Waddell TK, Hwang DM, Liu M, Keshavjee S
    Mol. Ther., 212;20(6):1204-11.
    Species: Porcine
    Sample Types: Plasma

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Porcine TNF-alpha Quantikine ELISA Kit
By Anonymous on 12/01/2017
Sample Tested: Porcine Serum