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Mouse TNF-alpha Antibody

Catalog #: MAB4101
35 Citations Datasheet
Catalog # Availability Size / Price Qty
MAB4101
MAB4101-SP
Detection of TNF-alpha in Raw264 Mouse Cell Line by Flow Cytometry.
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Citations (35)
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Mouse TNF-alpha Antibody Summary

Species Reactivity
Mouse
Specificity
Detects mouse TNF-alpha.
Source
Monoclonal Rat IgG1 Clone # MP6-XT22
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
mouse TNF-alpha
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or 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
Intracellular Staining by Flow Cytometry
0.25 µg/106 cells
Raw264 cells treated with LPS, fixed and permeabilized with FlowX FoxP3/Transcription Factor Fixation & Perm Kit (Catalog # FC012)
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 0.15-0.75 µg/mL in the presence of 0.25 ng/mL Recombinant Mouse TNF‑ alpha and 1 µg/mL actinomycin D.

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.

Scientific Data

Intracellular Staining by Flow Cytometry View Larger

Detection of TNF-alpha in Raw264 Mouse Cell Line by Flow Cytometry. Raw264 mouse macrophage cell line treated with 1 μg/mL LPS overnight was stained with Rat Anti-Mouse TNF-alpha Monoclonal Antibody (Catalog # MAB4101, filled histogram) or isotype control antibody (MAB005, open histogram) followed by Anti-Rat IgG CFS-conjugated Secondary Antibody (F0104). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3/Transcription Factor Fixation & Perm Kit (FC012). Staining was performed using our Staining Intracellular Molecules protocol.

Intracellular Staining by Flow Cytometry View Larger

Detection of TNF-alpha in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes (A) treated with PMA (50 ng/mL), Ca2+ Ionomycin (200 ng/mL) and Brefeldin A (5 μg/mL) for 4 hours or (B) untreated, were stained with Rat Anti-Mouse TNF-alpha Monoclonal Antibody (Catalog # MAB4101) followed by Anti-Rat IgG PE-conjugated Secondary Antibody (F0105B) and Rat anti-Mouse CD3 APC-conjugated Monoclonal Antibody (FAB4841A). Quadrant markers were set based on isotype control antibody (MAB005). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (FC005). Staining was performed using our protocol for Staining Intracellular Molecules.

Neutralization Cytotoxicity Induced by TNF‑ alpha and Neutralization by Mouse TNF‑ alpha Antibody. View Larger

Cytotoxicity Induced by TNF‑ alpha and Neutralization by Mouse TNF‑ alpha Antibody. Recombinant Mouse TNF-a (Catalog # 410-MT) induces cytotoxicity in the the L-929 mouse fibroblast cell line in a dose-dependent manner (orange line). Cytotoxicity elicited by Recombinant Mouse TNF-a (0.25 ng/mL) is neutralized (green line) by increasing concentrations of Mouse TNF-a Monoclonal Antibody (Catalog # MAB4101). The ND50 is typically 0.15-0.75 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).

Western Blot Detection of Mouse TNF-alpha by Western Blot View Larger

Detection of Mouse TNF-alpha by Western Blot Parameters of inflammation/nitrosative stress are unaltered in adipose tissue after chronic administration of compound 21. (A) and (B) Levels of TNF‐ alpha determined by WB and IHC. (C) and (D) levels of nitrotyrosine determined by WB and IHC. Quantification of specific bands was performed with Gel‐Pro Analyzer software. (C) For nitrotyrosine quantification, the intensity of the most predominant bands in the WB (of unknown identity), with MWs of 53 and 65 kDa, respectively, were quantified in each sample. Bar graphs are the means ± SE. (B) and (D) Positive TNF‐ alpha and nitrotyrosine staining as detected by immunohistochemistry was quantified using Image‐Pro Plus software. Data were analyzed by unpaired two‐tailed Student's t test. C21, n = 9 per group, control, n = 12 per group for TNF‐ alpha quantification. For nitrotyrosine quantification by Western blotting: C21, n = 6 per group; control, n = 6 per group and by immunohistochemistry: C21, n = 9 per group; control, n = 9 per group. WB, western blotting. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30156060), licensed under a CC-BY license. Not internally tested by R&D Systems.

Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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, TNF- alpha, TNFA ), 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, immune system development, apoptosis, and lipid metabolism. TNF-alpha is produced by several lymphoid cells as well as by astrocytes, endothelial cells, and smooth muscle cells. Mouse TNF-alpha consists of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 179 aa extracellular domain (ECD). Within the ECD, mouse TNF-alpha shares 94% aa sequence identity with rat and 70%-77% with bovine, canine, cotton rat, equine, feline, human, porcine, and rhesus TNF-alpha. TNF-alpha is produced by a wide variety of immune, epithelial, endothelial, and tumor cells. TNF-alpha is assembled intracellularly to form a noncovalently linked homotrimer which is expressed on the cell surface. Cell surface TNF-alpha can induce the lysis of neighboring tumor cells and virus infected cells, and it can generate its own downstream cell signaling following ligation by soluble TNFR I. Shedding of membrane bound TNF-alpha by TACE/ADAM17 releases the bioactive cytokine, a 55 kDa molecular weight soluble trimer of the TNF-alpha extracellular domain. TNF-alpha binds the ubiquitous 55-60 kDa TNF RI and the hematopoietic cell-restricted 80 kDa TNF RII, both of which are also expressed as homotrimers present on virtually all cell types. Both type I and type II receptors bind TNF-alpha with comparable affinity, although only TNF RI contains a cytoplasmic death domain which triggers the activation of apoptosis. Soluble forms of both types of receptors are released and can neutralize the biological activity of TNF-alpha.

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

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Citations for Mouse TNF-alpha Antibody

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.

35 Citations: Showing 1 - 10
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  1. Targeting WDxR motif reprograms immune microenvironment and inhibits hepatocellular carcinoma progression
    Authors: H Zhang, G Chen, X Feng, H Song, L Meng, Y Fu, J Yang, Z Fan, Y Ding, Z Du, J Wang, L Yang, J Zhang, L Sun, Z Liu, Z Zhang, Q Li, X Fan
    Embo Molecular Medicine, 2023-03-22;0(0):e15924.
  2. Functionalized nanowires for miRNA-mediated therapeutic programming of naïve T cells
    Authors: Yee Mon, KJ;Kim, S;Dai, Z;West, JD;Zhu, H;Jain, R;Grimson, A;Rudd, BD;Singh, A;
    Nature nanotechnology
    Species: Transgenic Mouse
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  3. Embryonic vitamin D deficiency programs hematopoietic stem cells to induce type 2 diabetes
    Authors: Oh, J;Riek, AE;Bauerle, KT;Dusso, A;McNerney, KP;Barve, RA;Darwech, I;Sprague, JE;Moynihan, C;Zhang, RM;Kutz, G;Wang, T;Xing, X;Li, D;Mrad, M;Wigge, NM;Castelblanco, E;Collin, A;Bambouskova, M;Head, RD;Sands, MS;Bernal-Mizrachi, C;
    Nature communications
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Neutralization
  4. Severe influenza infection is associated with inflammatory programmed cell death in infected macrophages
    Authors: André C. Ferreira, Carolina Q. Sacramento, Filipe S. Pereira-Dutra, Natália Fintelman-Rodrigues, Priscila P. Silva, Mayara Mattos et al.
    Frontiers in Cellular and Infection Microbiology
  5. EST12 regulates Myc expression and enhances anti-mycobacterial inflammatory response via RACK1-JNK-AP1-Myc immune pathway
    Authors: Jian Wu, Feng-Ling Luo, Yan Xie, Huan Xiong, Yadong Gao, Guanghui Liu et al.
    Frontiers in Immunology
  6. Activating a collaborative innate-adaptive immune response to control metastasis
    Authors: Lijuan Sun, Tim Kees, Ana Santos Almeida, Bodu Liu, Xue-Yan He, David Ng et al.
    Cancer Cell
  7. ADAM17 Boosts Cholesterol Efflux and Downstream Effects of High-Density Lipoprotein on Inflammatory Pathways in Macrophages
    Authors: Vishal Kothari, Jingjing Tang, Yi He, Farah Kramer, Jenny E. Kanter, Karin E. Bornfeldt
    Arteriosclerosis, Thrombosis, and Vascular Biology
  8. A class of viral inducer of degradation of the necroptosis adaptor RIPK3 regulates virus-induced inflammation
    Authors: Zhijun Liu, Himani Nailwal, Jonah Rector, Masmudur M. Rahman, Richard Sam, Grant McFadden et al.
    Immunity
  9. ABIN-1 is a key regulator in RIPK1-dependent apoptosis (RDA) and necroptosis, and ABIN-1 deficiency potentiates necroptosis-based cancer therapy in colorectal cancer
    Authors: J Cai, D Hu, J Sakya, T Sun, D Wang, L Wang, X Mao, Z Su
    Cell Death & Disease, 2021-02-01;12(2):140.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: IP
  10. Spinal interleukin-6 contributes to central sensitisation and persistent pain hypersensitivity in a model of juvenile idiopathic arthritis
    Authors: Charlie H.T. Kwok, Annastazia E Learoyd, Julia Canet-Pons, Tuan Trang, Maria Fitzgerald
    Brain, Behavior, and Immunity
  11. Myeloma Cells Down‐Regulate Adiponectin in Bone Marrow Adipocytes Via TNF‐Alpha
    Authors: Emma V Morris, Karla J Suchacki, Joseph Hocking, Rachel Cartwright, Aneka Sowman, Beatriz Gamez et al.
    Journal of Bone and Mineral Research
  12. Inflammation induced by incomplete radiofrequency ablation accelerates tumor progression and hinders PD-1 immunotherapy
    Authors: L Shi, J Wang, N Ding, Y Zhang, Y Zhu, S Dong, X Wang, C Peng, C Zhou, L Zhou, X Li, H Shi, W Wu, X Long, C Wu, W Liao
    Nat Commun, 2019-11-28;10(1):5421.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Cell Culture
  13. A Bcr-Abl Inhibitor GNF-2 Attenuates Inflammatory Activation of Glia and Chronic Pain
    Authors: GJ Song, MH Rahman, MK Jha, DP Gupta, SH Park, JH Kim, SH Lee, IK Lee, T Sim, YC Bae, WH Lee, K Suk
    Front Pharmacol, 2019-05-20;10(0):543.
    Species: Mouse
    Sample Types: Cell Culture Supernates
    Applications: ELISA Capture
  14. Neuregulin-1 Fosters Supportive Interactions between Microglia and Neural Stem/Progenitor Cells
    Authors: Ghazaleh M. Shahriary, Hardeep Kataria, Soheila Karimi-Abdolrezaee
    Stem Cells International
  15. Iron alters macrophage polarization status and leads to steatohepatitis and fibrogenesis
    Authors: P Handa, S Thomas, V Morgan-Ste, BD Maliken, E Gochanour, S Boukhar, MM Yeh, KV Kowdley
    J. Leukoc. Biol., 2019-03-05;0(0):.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  16. Regulation of the bi-directional cross-talk between ovarian cancer cells and adipocytes by SPARC
    Authors: B John, C Naczki, C Patel, A Ghoneum, S Qasem, Z Salih, N Said
    Oncogene, 2019-02-14;0(0):.
    Species: Mouse
    Sample Types: In Vivo
    Applications: Neutralization
  17. Monosodium urate crystals reduce osteocyte viability and indirectly promote a shift in osteocyte function towards a proinflammatory and proresorptive state
    Authors: A Chhana, B Pool, KE Callon, ML Tay, D Musson, D Naot, G McCarthy, S McGlashan, J Cornish, N Dalbeth
    Arthritis Res. Ther., 2018-09-10;20(1):208.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Neutralization
  18. Chronic administration of the angiotensin type 2 receptor agonist C21 improves insulin sensitivity in C57BL/6 mice
    Authors: DT Quiroga, MC Muñoz, C Gil, M Pffeifer, JE Toblli, UM Steckeling, JF Giani, FP Dominici
    Physiol Rep, 2018-08-01;6(16):e13824.
    Species: Mouse
    Sample Types: Tissue Homogenates, Whole Tissue
    Applications: IHC, Western Blot
  19. TNF and granulocyte macrophage-colony stimulating factor interdependence mediates inflammation via CCL17
    Authors: AD Cook, MC Lee, R Saleh, HW Khiew, AD Christense, A Achuthan, AJ Fleetwood, DC Lacey, JE Smith, I Förster, JA Hamilton
    JCI Insight, 2018-03-22;3(6):.
    Species: Mouse
    Sample Types: In Vivo
    Applications: Neutralization
  20. Melatonin induces anti-inflammatory effects via endoplasmic reticulum stress in RAW264.7 macrophages
    Authors: Yina Chen, Qian Zhao, Yangjie Sun, Yin Jin, Jie Zhang, Jiansheng Wu
    Molecular Medicine Reports
  21. Helicobacter pylori Secreted Protein HP1286 Triggers Apoptosis in Macrophages via TNF-Independent and ERK MAPK-Dependent Pathways
    Authors: Raquel Tavares, Sushil Kumar Pathak
    Frontiers in Cellular and Infection Microbiology
  22. Induction of necroptotic cell death by viral activation of the RIG-I or STING pathway
    Authors: SN Schock, NV Chandra, Y Sun, T Irie, Y Kitagawa, B Gotoh, L Coscoy, A Winoto
    Cell Death Differ, 2017-01-06;0(0):.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  23. Presentation of Cryptic Peptides by MHC Class I Is Enhanced by Inflammatory Stimuli
    J Immunol, 2016-09-19;0(0):.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Neutralization
  24. Myostatin is a direct regulator of osteoclast differentiation and its inhibition reduces inflammatory joint destruction in mice.
    Authors: Dankbar B, Fennen M, Brunert D, Hayer S, Frank S, Wehmeyer C, Beckmann D, Paruzel P, Bertrand J, Redlich K, Koers-Wunrau C, Stratis A, Korb-Pap A, Pap T
    Nat Med, 2015-08-03;21(9):1085-90.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Neutralization
  25. Microglia-induced IL-6 protects against neuronal loss following HSV-1 infection of neural progenitor cells
    Authors: Ana J. Chucair-Elliott, Christopher Conrady, Min Zheng, Chandra M. Kroll, Thomas E. Lane, Daniel J. J. Carr
    Glia
  26. MyD88 signalling in myeloid cells is sufficient to prevent chronic mycobacterial infection.
    Authors: Berod L, Stuve P, Swallow M, Arnold-Schrauf C, Kruse F, Gentilini M, Freitag J, Holzmann B, Sparwasser T
    Eur J Immunol, 2014-02-13;44(5):1399-409.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  27. A paracrine network regulates the cross-talk between human lung stem cells and the stroma.
    Authors: Ruiz E, Oeztuerk-Winder F, Ventura J
    Nat Commun, 2014-01-01;5(0):3175.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  28. Acute lung injury and acute kidney injury are established by four hours in experimental sepsis and are improved with pre, but not post, sepsis administration of TNF-alpha antibodies.
    Authors: Bhargava R, Altmann C, Andres-Hernando A, Webb R, Okamura K, Yang Y, Falk S, Schmidt E, Faubel S
    PLoS ONE, 2013-11-12;8(11):e79037.
    Species: Mouse
    Sample Types: In Vivo
    Applications: Neutralization
  29. Helicobacter pylori Protein JHP0290 Binds to Multiple Cell Types and Induces Macrophage Apoptosis via Tumor Necrosis Factor (TNF)-Dependent and Independent Pathways
    Authors: Sushil Kumar Pathak, Raquel Tavares, Nele de Klerk, Anna-Lena Spetz, Ann-Beth Jonsson
    PLoS ONE
  30. Critical involvement of extracellular ATP acting on P2RX7 purinergic receptors in photoreceptor cell death.
    Authors: Notomi S, Hisatomi T, Kanemaru T, Takeda A, Ikeda Y, Enaida H, Kroemer G, Ishibashi T
    Am. J. Pathol., 2011-10-08;179(6):2798-809.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Neutralization
  31. Toll-like receptor 3 ligand dampens liver inflammation by stimulating Valpha 14 invariant natural killer T cells to negatively regulate gammadeltaT cells.
    Authors: Gardner TR, Chen Q, Jin Y, Ajuebor MN
    Am. J. Pathol., 2010-02-18;176(4):1779-89.
    Species: Mouse
    Sample Types: In Vivo
    Applications: Neutralization
  32. Relative importance of T-cell subsets in monocytotropic ehrlichiosis: a novel effector mechanism involved in Ehrlichia-induced immunopathology in murine ehrlichiosis.
    Authors: Ismail N, Crossley EC, Stevenson HL, Walker DH
    Infect. Immun., 2007-06-11;75(9):4608-20.
    Species: Mouse
    Sample Types: In Vivo
    Applications: Neutralization
  33. CCL16/LEC powerfully triggers effector and antigen-presenting functions of macrophages and enhances T cell cytotoxicity.
    Authors: Cappello P, Caorsi C, Bosticardo M
    J. Leukoc. Biol., 2003-10-02;75(1):135-42.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Neutralization
  34. Macrophages activated by hepatitis B virus have distinct metabolic profiles and suppress the virus via IL-1beta to downregulate PPARalpha and FOXO3
    Authors: Y Li, Y Zhu, S Feng, Y Ishida, TP Chiu, T Saito, S Wang, DK Ann, JJ Ou
    Cell Reports, 2022-01-25;38(4):110284.

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