Human TNF-alpha Antibody

(18 citations)
(1 Review)
  
  • Species Reactivity
    Human
  • Specificity
    Detects human TNF‑ alpha in direct ELISAs and Western blots. In direct ELISAs, approximately 50% cross‑reactivity with recombinant feline TNF‑ alpha, recombinant canine TNF‑ alpha, recombinant rhesus monkey TNF‑ alpha, and recombinant equine TNF-alpha is observed and approximately 15% cross‑reactivity with recombinant cotton rat TNF‑ alpha, recombinant mouse TNF‑ alpha, recombinant rat TNF‑ alpha, and recombinant guinea pig TNF‑ alpha  is observed and less than 5% cross‑reactivity with recombinant porcine TNF‑ alpha and recombinant bovine TNF‑ alpha  is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human TNF-alpha
    Val77-Leu233
    Accession # P01375
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    Recombinant Human TNF‑ alpha (Catalog # 210-TA)
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize TNF‑ alpha -induced cytotoxicity in the L‑929 mouse fibroblast cell line. Matthews, N. and M. L. Neale (1987) in Lymphokines and Interferons, A Practical Approach. Clemens, M. J. et al. (eds): IRL Press. 221. The Neutralization Dose (ND50) is typically 0.01-0.06 μg/ml in the presence of 0.75 ng/mL Recombinant Human TNF‑ alpha.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Cytotoxicity Induced by TNF‑ alpha and Neutralization by Human TNF-alpha Antibody.

Recombinant Human TNF‑ alpha (Catalog # 210-TA) induces cytotoxicity in the L‑929 mouse fibroblast cell line in a dose-dependent manner (orange line). Cytotoxicity elicited by Recombinant Human TNF‑ alpha (0.75 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human TNF‑ alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-210-NA). The ND50 is typically 0.01‑0.06 μg/mL.

Immunocytochemistry
TNF‑ alpha in Human PBMCs. TNF‑ alpha was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Goat Anti-Human TNF‑ alpha Antigen Affinity-purified Polyclonal Antibody (Catalog #
AF-210-NA) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: TNF-alpha
Tumor necrosis factor alpha (TNF-alpha ), 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-alpha is produced by a wide variety of immune and epithelial cell types. Human TNF-alpha 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-alpha shares 97% aa sequence identity with rhesus and 71%‑92% with bovine, canine, cotton rat, equine, feline, mouse, porcine, and rat TNF-alpha. 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-alpha by TACE/ADAM17 releases a 55 kDa soluble trimeric form of TNF-alpha. TNF-alpha trimers bind the ubiquitous TNF RI and the hematopoietic cell-restricted TNF RII, both of which are also expressed as homotrimers. TNF-alpha regulates lymphoid tissue development through control of apoptosis. It also promotes inflammatory responses by inducing the activation of vascular endothelial cells and macrophages. TNF-alpha 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
Related Research Areas
Citations:

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.

18 Citations: Showing 1 - 10
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Species
Applications
Sample Type
  1. Inflammatory Adipokines Decrease Expression of Two High Molecular Weight Isoforms of Tropomyosin Similar to the Change in Type 2 Diabetic Patients
    PLoS ONE, 2016;11(9):e0162908.
    Species: Human
    Sample Type: Whole Cells
    Application: Bioassay
  2. Licensed human natural killer cells aid dendritic cell maturation via TNFSF14/LIGHT.
    Authors: Holmes T, Wilson E, Black E, Benest A, Vaz C, Tan B, Tanavde V, Cook G
    Proc Natl Acad Sci U S A, 2014;111(52):E5688-96.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  3. Reference range and short- and long-term biological variation of interleukin (IL)-6, IL-17A and tissue necrosis factor-alpha using high sensitivity assays.
    Authors: Todd J, Simpson P, Estis J, Torres V, Wub A
    Cytokine, 2013;64(3):660-5.
    Species: Human
    Sample Type: Plasma
    Application: ELISA Development
  4. The tachykinins substance P and hemokinin-1 favor the generation of human memory Th17 cells by inducing IL-1beta, IL-23, and TNF-like 1A expression by monocytes.
    Authors: Cunin P, Caillon A, Corvaisier M, Garo E, Scotet M, Blanchard S, Delneste Y, Jeannin P
    J. Immunol., 2011;186(7):4175-82.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  5. Native IL-32 is released from intestinal epithelial cells via a non-classical secretory pathway as a membrane-associated protein.
    Authors: Hasegawa H, Thomas HJ, Schooley K, Born TL
    Cytokine, 2011;53(1):74-83.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  6. Thymic stromal lymphopoietin from trophoblasts induces dendritic cell-mediated regulatory TH2 bias in the decidua during early gestation in humans.
    Authors: Guo PF, Du MR, Wu HX, Lin Y, Jin LP, Li DJ
    Blood, 2010;116(12):2061-9.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC - Not specified
  7. Disordered macrophage cytokine secretion underlies impaired acute inflammation and bacterial clearance in Crohn's disease.
    Authors: Smith AM, Rahman FZ, Hayee B, Graham SJ, Marks DJ, Sewell GW, Palmer CD, Wilde J, Foxwell BM, Gloger IS, Sweeting T, Marsh M, Walker AP, Bloom SL, Segal AW
    J. Exp. Med., 2009;206(9):1883-97.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  8. KiSS1 metastasis suppressor gene product induces suppression of tyrosine kinase receptor signaling to Akt, tumor necrosis factor family ligand expression, and apoptosis.
    Authors: Navenot JM, Fujii N, Peiper SC
    Mol. Pharmacol., 2009;75(5):1074-83.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  9. Bronchial epithelial cell growth regulation in fibroblast cocultures: the role of hepatocyte growth factor.
    Authors: Skibinski G, Elborn JS, Ennis M
    Am. J. Physiol. Lung Cell Mol. Physiol., 2007;293(1):L69-76.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  10. Simultaneous induction of apoptotic and survival signaling pathways in macrophage-like THP-1 cells by Shiga toxin 1.
    Authors: Lee SY, Cherla RP, Tesh VL
    Infect. Immun., 2007;75(3):1291-302.
    Species: Human
    Sample Type: Whole Cells
    Application: Functional Assay
  11. Spatial compartmentalization of tumor necrosis factor (TNF) receptor 1-dependent signaling pathways in human airway smooth muscle cells. Lipid rafts are essential for TNF-alpha-mediated activation of RhoA but dispensable for the activation of the NF-kappaB and MAPK pathways.
    Authors: Hunter I, Nixon GF
    J. Biol. Chem., 2006;281(45):34705-15.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  12. Sepsis and pathophysiology of anthrax in a nonhuman primate model.
    Authors: Stearns-Kurosawa DJ, Lupu F, Taylor FB, Kinasewitz G, Kurosawa S
    Am. J. Pathol., 2006;169(2):433-44.
    Species: Primate - Papio cynocephalus (Yellow Baboon)
    Sample Type: Plasma
    Application: ELISA Development
  13. Receptors and lytic mediators regulating anti-tumor activity by the leukemic killer T cell line TALL-104.
    Authors: Brando C, Mukhopadhyay S, Kovacs E, Medina R, Patel P, Catina TL, Campbell KS, Santoli D
    J. Leukoc. Biol., 2005;78(2):359-71.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  14. Tumor necrosis factor, interleukin 11, and leukemia inhibitory factor produced by Langerhans cells in Langerhans cell histiocytosis.
    Authors: Andersson By U, Tani E, Andersson U
    J. Pediatr. Hematol. Oncol., 2004;26(11):706-11.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  15. Influence of eicosapentaenoic acid, an omega-3 fatty acid, on ultraviolet-B generation of prostaglandin-E2 and proinflammatory cytokines interleukin-1 beta, tumor necrosis factor-alpha, interleukin-6 and interleukin-8 in human skin in vivo.
    Authors: Shahbakhti H, Watson RE, Azurdia RM, Ferreira CZ, Garmyn M, Rhodes LE
    Photochem. Photobiol., 2004;80(2):231-5.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC
  16. Interleukin-8 production in tuberculous pleurisy: role of mesothelial cells stimulated by cytokine network involving tumour necrosis factor-alpha and interleukin-1 beta.
    Authors: Park JS, Kim YS, Jee YK, Myong NH, Lee KY
    Scand. J. Immunol., 2003;57(5):463-9.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  17. Cytotoxicity and interleukin-1beta processing following Shigella flexneri infection of human monocyte-derived dendritic cells.
    Authors: Edgeworth JD, Spencer J, Phalipon A, Griffin GE, Sansonetti PJ
    Eur. J. Immunol., 2002;32(5):1464-71.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  18. Natural killer cell-produced IFN-gamma and TNF-alpha induce target cell cytolysis through up-regulation of ICAM-1.
    Authors: Wang R, Jaw JJ, Jaw J, Stutzman N, Stutzman NC, Zou Z, Sun PD
    J Leukoc Biol, 0;91(2):299-309.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
Expand to show all 18 Citations
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Average Rating: 5 (Based on 1 review)

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ELISA Human TNF-alpha  Antibody AF-210-NA
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 ELISA Human Anonymous 11/16/2017
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ELISA Human TNF-alpha  Antibody AF-210-NA
ELISA: Human TNF-alpha Antibody [AF-210-NA]

Summary

ApplicationELISA
Sample TestedSerum and Plasma
SpeciesHuman

Other Experimental Details

Other Experimental DetailsThis antibody was used with MAB610 to build an ELISA to measure TNFa. This antibody was used as the detection while MAB610 was used as the capture.

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