Human TNF-alpha Biotinylated Antibody

(22 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human TNF-alpha in ELISAs and Western blots. In sandwich immunoassays, less than 0.04% cross-reactivity with recombinant human (rh) TNF‑ beta, recombinant mouse (rm) TNF-alpha, recombinant rat TNF‑ alpha, recombinant porcine TNF-alpha, rhsTNF RII, rmsTNF RI, and rmsTNF RII is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human TNF-alpha (R&D Systems, Catalog # 210-TA)
    Val77-Leu233
    Accession # P01375
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
  • Label
    Biotin
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    Recombinant Human TNF‑ alpha (Catalog # 210-TA)
  • Immunocytochemistry
    5-15 µg/mL
    See below
    • Human TNF-alpha Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    2-8 µg/mL 
    Human TNF‑ alpha Antibody (Catalog # MAB610R)
  • ELISA Capture (Matched Antibody Pair)
    2-8 µg/mL 
    Human TNF‑ alpha Antibody (Catalog # MAB610)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Human TNF‑ alpha Biotinylated Antibody (Catalog # BAF210)
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
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 Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF210) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Streptavidin (red; Catalog # NL999) 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.
  • 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 (1, 2). Human TNF-alpha consists of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD) (3). 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 (4). Ligation of this complex induces reverse signaling that promotes lymphocyte costimulation but diminishes monocyte responsiveness (5). Cleavage of membrane bound TNF-alpha by TACE/ADAM17 releases a 55 kDa soluble trimeric form of TNF-alpha (6, 7). TNF-alpha trimers bind the ubiquitous TNF RI and the hematopoietic cell-restricted TNF RII, both of which are also expressed as homotrimers (1, 8). TNF-alpha regulates lymphoid tissue development through control of apoptosis (2). It also promotes inflammatory responses by inducing the activation of vascular endothelial cells and macrophages (2). TNF-alpha is a key cytokine in the development of several inflammatory disorders (9). It contributes to the development of type 2 diabetes through its effects on insulin resistance and fatty acid metabolism (10, 11).

  • References:
    1. Idriss, H.T. and J.H. Naismith (2000) Microsc. Res. Tech. 50:184.
    2. Hehlgans, T. and K. Pfeffer (2005) Immunology 115:1.
    3. Pennica, D. et al. (1984) Nature 312:724.
    4. Tang, P. et al. (1996) Biochemistry 35:8216.
    5. Eissner G. et al. (2004) Cytokine Growth Factor Rev. 15:353.
    6. Black, R.A. et al. (1997) Nature 385:729.
    7. Moss, M.L. et al. (1997) Nature 385:733.
    8. Loetscher, H. et al. (1991) J. Biol. Chem. 266:18324.
    9. Clark, I.A. (2007) Cytokine Growth Factor Rev. 18:335.
    10. Romanatto, T. et al. (2007) Peptides 28:1050.
    11. Hector, J. et al. (2007) Horm. Metab. Res. 39:250.
  • 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.

22 Citations: Showing 1 - 10
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Species
Applications
Sample Type
  1. Interleukin-10 and prostaglandin E2 have complementary but distinct suppressive effects on Toll-like receptor-mediated dendritic cell activation in ovarian carcinoma
    Authors: E Brencicova, AL Jagger, HG Evans, M Georgouli, A Laios, S Attard Mon, G Mehra, J Spencer, AA Ahmed, S Raju-Kanki, LS Taams, SS Diebold
    PLoS ONE, 2017;12(4):e0175712.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: ELISA detection
  2. Porphyromonas gingivalis gingipains cause defective macrophage migration towards apoptotic cells and inhibit phagocytosis of primary apoptotic neutrophils
    Authors: SA Castro, R Collighan, PA Lambert, IH Dias, P Chauhan, CE Bland, I Milic, MR Milward, PR Cooper, A Devitt
    Cell Death Dis, 2017;8(3):e2644.
    Species: Human
    Sample Type: Cell Culture Supernates
  3. Elevated Membrane and Soluble CD64: A Novel Marker Reflecting Altered FcgammaR Function and Disease in Early Rheumatoid Arthritis That Can Be Regulated by Anti-Rheumatic Treatment.
    Authors: Matt P, Lindqvist U, Kleinau S
    PLoS ONE, 2015;10(9):e0137474.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: ELISA detection
  4. C-reactive protein promotes diabetic kidney disease in a mouse model of type 1 diabetes.
    Authors: Liu F, Chen HY, Huang XR, Chung AC, Zhou L, Fu P, Szalai AJ, Lan HY
    Diabetologia, 2011;54(10):2713-23.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: ELISA Development
  5. Protection from RNA and DNA viruses by IL-32.
    Authors: Zepp JA, Nold-Petry CA, Dinarello CA, Nold MF
    J. Immunol., 2011;186(7):4110-8.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: Electrochemiluminescence
  6. Resistance of human alveolar macrophages to Bacillus anthracis lethal toxin.
    Authors: Wu W, Mehta H, Chakrabarty K, Booth JL, Duggan ES, Patel KB, Ballard JD, Coggeshall KM, Metcalf JP
    J. Immunol., 2009;183(9):5799-806.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: ELISA Development
  7. Noninvasive detection of acute and chronic injuries in human renal transplant by elevation of multiple cytokines/chemokines in urine.
    Authors: Hu H, Kwun J, Aizenstein BD, Knechtle SJ
    Transplantation, 2009;87(12):1814-20.
    Species: Human
    Sample Type: Urine
    Application: Antibody Array Development
  8. A novel and sensitive ELISA reveals that the soluble form of IL-13R-alpha2 is not expressed in plasma of healthy or asthmatic subjects.
    Authors: O'Toole M, Legault H, Ramsey R, Wynn TA, Kasaian MT
    Clin. Exp. Allergy, 2008;38(4):594-601.
    Species: Human
    Sample Type: BALF
    Application: ELISA Development
  9. Lipoteichoic acid isolated from Lactobacillus plantarum inhibits lipopolysaccharide-induced TNF-alpha production in THP-1 cells and endotoxin shock in mice.
    Authors: Kim HG, Kim NR, Gim MG, Lee JM, Lee SY, Ko MY, Kim JY, Han SH, Chung DK
    J. Immunol., 2008;180(4):2553-61.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: ELISA Development
  10. Development and validation of sandwich ELISA microarrays with minimal assay interference.
    Authors: Gonzalez RM, Seurynck-Servoss SL, Crowley SA
    J. Proteome Res., 2008;7(6):2406-14.
    Species: Human
    Sample Type: Serum
    Application: ELISA Microarray Development
  11. Upregulation of human cytomegalovirus by HIV type 1 in human lymphoid tissue ex vivo.
    Authors: Biancotto A, Iglehart SJ, Lisco A, Vanpouille C, Grivel JC, Lurain NS, Reichelderfer PS, Margolis LB
    AIDS Res. Hum. Retroviruses, 2008;24(3):453-62.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: Luminex Development
  12. Ultrasensitive flow-based immunoassays using single-molecule counting.
    Authors: Todd J, Freese B, Lu A, Held D, Morey J, Livingston R, Goix P
    Clin. Chem., 2007;53(11):1990-5.
    Species: Human
    Sample Type: Plasma
    Application: ELISA Development
  13. Abnormal activation and cytokine spectra in lymph nodes of people chronically infected with HIV-1.
    Authors: Biancotto A, Grivel JC, Iglehart SJ, Vanpouille C, Lisco A, Sieg SF, Debernardo R, Garate K, Rodriguez B, Margolis LB, Lederman MM
    Blood, 2007;109(10):4272-9.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: Luminex Development
  14. HIV-1 pathogenesis differs in rectosigmoid and tonsillar tissues infected ex vivo with CCR5- and CXCR4-tropic HIV-1.
    Authors: Grivel JC, Elliott J, Lisco A, Biancotto A, Condack C, Shattock RJ, McGowan I, Margolis L, Anton P
    AIDS, 2007;21(10):1263-72.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: Luminex Development
  15. Fluorescence single-molecule counting assays for high-sensitivity detection of cytokines and chemokines.
    Authors: Qui H, Ferrell EP, Nolan N, Phelps BH, Tabibiazar R, Whitney DH, Naelfski EA
    Clin. Chem., 2007;53(11):2010-2.
    Species: Human
    Sample Type: Plasma
    Application: ELISA Development
  16. Increased ENA-78 in the follicular fluid of patients with endometriosis.
    Authors: Wunder DM, Mueller MD, Birkhauser MH, Bersinger NA
    Acta Obstet Gynecol Scand, 2006;85(3):336-42.
    Species: Human
    Sample Type: Follicular Fluid
    Application: ELISA Development
  17. Selective regulation of IL-10 signaling and function by zymosan.
    Authors: Du Z, Kelly E, Mecklenbrauker I, Agle L, Herrero C, Paik P, Ivashkiv LB
    J. Immunol., 2006;176(8):4785-92.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: ELISA Development
  18. 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
  19. Direct comparison of traditional ELISAs and membrane protein arrays for detection and quantification of human cytokines.
    Authors: Copeland S, Siddiqui J, Remick D
    J. Immunol. Methods, 2004;284(1):99-106.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: Array Development
  20. trans fatty acids and systemic inflammation in heart failure.
    Authors: Mozaffarian D, Rimm EB, King IB, Lawler RL, McDonald GB, Levy WC
    Am. J. Clin. Nutr., 2004;80(6):1521-5.
    Species: Human
    Sample Type: Plasma
    Application: ELISA Development
  21. IFN-alpha priming results in a gain of proinflammatory function by IL-10: implications for systemic lupus erythematosus pathogenesis.
    Authors: Sharif MN, Tassiulas I, Hu Y, Mecklenbrauker I, Tarakhovsky A, Ivashkiv LB
    J. Immunol., 2004;172(10):6476-81.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: ELISA Development
  22. Reprogramming of IL-10 activity and signaling by IFN-gamma.
    Authors: Herrero C, Hu X, Li WP, Samuels S, Sharif MN, Kotenko S, Ivashkiv LB
    J. Immunol., 2003;171(10):5034-41.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: ELISA Development
Expand to show all 22 Citations
Isotype Controls
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