TNF-alpha Antibody

Western Blot: TNF-alpha Antibody - Azide Free [NB600-587] - Analysis using Azide/BSA FREE version of NB600-587. Membrane blocked in 1% BSA-TBS-T 30 min RT, 1:1000 dilution in 1% BSA-TBS-T 4C.

Western Blot: TNF-alpha Antibody - Azide Free [NB600-587] - Expression of TNF-alpha, IL-1beta and NF-kappaB in the brains of MCAO rats. Western blot confirmed that protein levels of TNF-alpha increased as early as 4 h after reperfusion, further increased at 24 h, peaked at 72 h and decreased at 7 days, respectively (*P<0.001, compared with the controls; #P<0.001 compared with the previous time point). Five rats in each group were used for western blot (n=5), sham, sham-operated control rats Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2014586), licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: TNF-alpha Antibody - Azide Free [NB600-587] - LMP2 knockdown by shRNA reduced the expression of TNF-alpha and IL-1beta after MCAO. Immunofluorescence labeling revealed that both TNF-alpha and IL-1beta immunoreactivity was significantly upregulated in the ipsilateral ischemic hemisphere compared with sham-operated animals. In contrast, LMP2 knockdown by shRNA significantly reduced the expression of TNF-alpha and IL-1beta. Five rats in each group were used for immunofluorescent assays (n=5). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2014586), licensed under a CC-BY license.

Immunohistochemistry: TNF-alpha Antibody - Azide Free [NB600-587] - Staining of formalin/PFA-fixed paraffin-embedded sections of human artery tissue sections. Sections were fixed in formaldehyde and subjected to heat mediated antigen retrieval in citrate buffer (pH 6.0). Slides were blocked for ten minutes with 1.5% serum. Primary antibody was diluted 1:100 and incubated with samples for 24 hours at 4C. HRP-conjugated goat anti-rabbit antibody was used as the secondary antibody.

Western Blot: TNF-alpha Antibody - Azide Free [NB600-587] - Expression of TNF-alpha, IL-1beta and NF-kappaB in the brains of MCAO rats. Western blot confirmed that protein levels of TNF-alpha increased as early as 4 h after reperfusion, further increased at 24 h, peaked at 72 h and decreased at 7 days, respectively (*P<0.001, compared with the controls; #P<0.001 compared with the previous time point). Five rats in each group were used for western blot (n=5), sham, sham-operated control rats Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2014586), licensed under a CC-BY license.

Fluorescent immunohistochemistry showing staining of human colon by anti-TNF alpha (formalin/PFA-fixed paraffin-embedded sections). Samples were formaldehyde-fixed, then blocked in 10% serum for 2 hours at 20C. The primary antibody was diluted 1:100 and incubated with the sample for 2 hours at 20C. Alexa Fluor(R) 680 goat polyclonal secondary antibody was used diluted 1:5000.

Immunohistochemistry using polyclonal TNFa antibody showing staining of formalin/PFA-fixed paraffin-embedded sections of human artery tissue sections. Sections were fixed in formaldehyde and subjected to heat mediated antigen retrieval in citrate buffer (pH 6.0). Slides were blocked for ten minutes with 1.5% serum. Primary antibody was diluted 1:100 and incubated with samples for 24 hours at 4C. HRP-conjugated goat anti-rabbit antibody was used as the secondary antibody.

Western blot of TNF-alpha Antibody - Azide Free. Lane 1: 250ng human recombinant TNF. Lane 2: 500ng human recombinant TNF. Lane 3: 1000ng human recombinant TNF. Membrane blocked in Blotto

• Reactivity: Hu, Mu, Rt, Ch, Pm
• Applications: WB, ELISA, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated

TNF-alpha Antibody Summary

• Description: Store vial at -20C prior to opening. Aliquot contents and freeze at -20C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.
• Immunogen: The whole rabbit serum was prepared by repeated immunizations with recombinant human TNF-alpha produced in E.coli. (Uniprot: P01375)
• Localization: Type 2 membrane protein or soluble form derived from the membrane by proteolytic cleavage.
• Specificity: The antiserum is directed against mature 17,000 MW human TNFa and is useful in determining its presence in various assays. In general, this antibody also detects primate TNFa in the same formats using similar dilutions. The antibody does not recognize human TNFb (lymphotoxin). This antiserum will recognize the cell-bound precursor of TNFa as a 26,000 protein in immunoblots, particularly in denatured samples. This antibody is also useful for neutralization of human and primate TNFa activity in bioassays. It does not neutralize the biological activity of lymphotoxin. For neutralization, it is recommended to incubate the sample with a 1:200 dilution of the antibody for at least 4 hours before being tested. A control of similarly diluted normal rabbit IgG is recommended.
• Isotype: Serum
• Clonality: Polyclonal
• Host: Rabbit
• Gene: TNF
• Purity: Whole Antiserum

Applications/Dilutions

• Dilutions:
  • ELISA 1:1000-1:5000
  • Immunocytochemistry/ Immunofluorescence 1:100
  • Immunohistochemistry 1:100-1:500
  • Immunohistochemistry-Paraffin 1:100-1:500
  • Western Blot 1:500-1:2000
• Application Notes: This product has been tested for use in immunohistochemistry, immunofluorescence, and immunoblotting. It recognizes the 17,000 MW TNFalpha. Reactivity in other immunoassays is unknown.

Reactivity Notes

Rat reactivity reported in scientific literature (PMID: 25894537), Mouse reactivity reported in scientific literature (PMID:22567116). Chicken reactivity reported in scientific literature (PMID: 30298003).

Packaging, Storage & Formulations

• Storage: Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.
• Buffer: Antiserum
• Preservative: No Preservative
• Purity: Whole Antiserum

Alternate Names for TNF-alpha Antibody

• 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

Background

Tumor necrosis factor (TNF)-alpha is a pro-inflammatory cytokine belonging to the TNF superfamily that is secreted by monocytes/macrophages, T cell, and natural killer (NK), among others (1). TNF-alpha is synthesized as a 233 amino acid (aa) transmembrane protein (mTNF-alpha) with a theoretical molecular weight (MW) of 26 kDa (1,2) that forms a homotrimer. mTNF-alpha is cleaved by TNF-alpha converting enzyme (TACE) and released in its 157 aa, 17 kDa soluble form (sTNF-alpha) (1-5). Both mTNF-alpha and sTNF-alpha are capable of binding type 1 TNF receptors (TNFR1), whereas mTNF-alpha predominately binds to TNFR2 (1,2). TNF-alpha binding to its receptors causes receptor recruitment of adaptor proteins, formation of signaling complexes, and downstream signaling cascades (e.g. MAPK, NF-kappaB, and Caspase-8), leading to distinct cellular responses such as survival, proliferation, inflammation, necroptosis, and apoptosis (1-5).

TNF-alpha is critical for normal immune response; however, dysregulation of TNF-alpha production can result in various pathologies (2,4,5). Excessive production of pro-inflammatory cytokines including interleukin 1 (IL-1), IL-6, and TNF-alpha has been implicated in an array of autoimmune diseases like rheumatoid arthritis (RA), inflammatory bowel disease (IBD), and psoriasis (2,4,5). Anti-TNF monoclonal antibodies, including Infliximab, and soluble TNFR have been approved for the treatment of autoimmune and TNF-mediated diseases (5). Additionally, data suggests that TNF inhibitors can be beneficial for treating patients experiencing immune-related adverse events associated with immune checkpoint inhibitor cancer treatment (6).

References

1. Holbrook J, Lara-Reyna S, Jarosz-Griffiths H, McDermott M. Tumour necrosis factor signalling in health and disease. F1000Res. 2019;8:F1000 Faculty Rev-111. https://doi.org/10.12688/f1000research.17023.1

2. Jang DI, Lee AH, Shin HY, et al. The Role of Tumor Necrosis Factor Alpha (TNF-alpha) in Autoimmune Disease and Current TNF-alpha Inhibitors in Therapeutics. Int J Mol Sci. 2021;22(5):2719. https://doi.org/10.3390/ijms22052719

3. Horiuchi T, Mitoma H, Harashima S, Tsukamoto H, Shimoda T. Transmembrane TNF-alpha: structure, function and interaction with anti-TNF agents. Rheumatology (Oxford). 2010;49(7):1215-1228. https://doi.org/10.1093/rheumatology/keq031

4. Webster JD, Vucic D. The Balance of TNF Mediated Pathways Regulates Inflammatory Cell Death Signaling in Healthy and Diseased Tissues. Front Cell Dev Biol. 2020;8:365. https://doi.org/10.3389/fcell.2020.00365

5. Kalliolias GD, Ivashkiv LB. TNF biology, pathogenic mechanisms and emerging therapeutic strategies. Nat Rev Rheumatol. 2016; 12(1):49-62. https://doi.org/10.1038/nrrheum.2015.169

6. Chen AY, Wolchok JD, Bass AR. TNF in the era of immune checkpoint inhibitors: friend or foe?. Nat Rev Rheumatol. 2021;17(4):213-223. doi:10.1038/s41584-021-00584-4

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for TNF-alpha Antibody (NB600-587)(31)

Publications using NB600-587

• Anna Flavia Cantone, Chiara Burgaletto, Giulia Di Benedetto, Anna Pannaccione, Agnese Secondo, Carlo Maria Bellanca, Egle Augello, Antonio Munafò, Paola Tarro, Renato Bernardini, Giuseppina Cantarella Taming Microglia in Alzheimer's Disease: Exploring Potential Implications of Choline Alphoscerate via alpha 7 nAChR Modulation. Cells 2024-02-26 [PMID: 38391922]
• T. Ollewagen, R.M. Benecke, C. Smith High species homology potentiates quantitative inflammation profiling in zebrafish using immunofluorescence Heliyon 2023-12-13 [PMID: 38187273]
• Ya‐Li Zhang, Ya‐Lei Wang, Ke Yan, Haiyan Li, Xinyu Zhang, Julien Milon Essola, Chengcheng Ding, Kexin Chang, Guangchao Qing, Fuxue Zhang, Yan Tan, Tiantian Peng, Xu Wang, Miao Jiang, Xing‐Jie Liang, Qian Hua Traditional Chinese Medicine Formulae QY305 Reducing Cutaneous Adverse Reaction and Diarrhea by its Nanostructure Advanced Science 2023-12-03 [PMID: 38044276]
• Corsetti G, Romano C, Pasini E et al. Essential Amino Acids-Rich Diet Increases Cardiomyocytes Protection in Doxorubicin-Treated Mice Nutrients 2023-05-12 [PMID: 37242170] (IHC, Mouse)
• Albadawy R, Hasanin AH, Agwa SHA et al. Rosavin Ameliorates Hepatic Inflammation and Fibrosis in the NASH Rat Model via Targeting Hepatic Cell Death International Journal of Molecular Sciences 2022-09-05 [PMID: 36077546] (Immunohistochemistry, Human)
• Kaya D, Micili SC, Kizmazoglu C et al. Allopurinol attenuates repeated traumatic brain injury in old rats: A preliminary report Experimental neurology 2022-08-02 [PMID: 35931122] (IHC-P, Rat)
• Omar AE, Al-Khalaifah HS, Osman A et al. Modulating the Growth, Antioxidant Activity, and Immunoexpression of Proinflammatory Cytokines and Apoptotic Proteins in Broiler Chickens by Adding Dietary Spirulina platensis Phycocyanin Antioxidants (Basel) 2022-05-19 [PMID: 35624855] (Immunohistochemistry, Human)
• Gutziet O, Iluz R, Ben Asher H et al. Maternal N-Acetyl-Cysteine Prevents Neonatal Hypoxia-Induced Brain Injury in a Rat Model International journal of molecular sciences 2021-12-20 [PMID: 34948425] (WB, Rat)
• Ollewagen T, Powrie YSL, Myburgh KH, Smith C Unresolved intramuscular inflammation, not diminished skeletal muscle regenerative capacity, is at the root of rheumatoid cachexia: insights from a rat CIA model Physiological reports 2021-11-01 [PMID: 34806343] (WB, Rat)
• Ongoz Dede F, Bozkurt Dogan S, Balli U Et al. The effect of ellagic acid on the repair process of periodontal defects related to experimental periodontitis in rats Journal of applied oral science : revista FOB 2021-09-27 [PMID: 34586188] (IF/IHC, Rat)
• Parvin S, Williams CR, Jarrett SA, Garraway SM Spinal Cord Injury Increases Pro-inflammatory Cytokine Expression in Kidney at Acute and Sub-chronic Stages Inflammation 2021-08-21 [PMID: 34417952] (WB, Mouse)
• Conti F, Romano GL, Eandi CM et al. Brimonidine is Neuroprotective in Animal Paradigm of Retinal Ganglion Cell Damage Frontiers in Pharmacology 2021-07-21 [PMID: 34366858] (Immunohistochemistry, Human)
• Corsetti G, Romano C, Pasini E et al. Qualitative Nitrogen Malnutrition Damages Gut and Alters Microbiome in Adult Mice. A Preliminary Histopathological Study Nutrients 2021-03-26 [PMID: 33810512] (IF/IHC, Mouse)
• Gomez-Lopez N, Romero R, Panaitescu B et al. Zinc gluconate-loaded chitosan nanoparticles reduce severity of collagen-induced arthritis in Wistar rats ACS Biomater Sci Eng 2021-01-06 [PMID: 33405580]
• Bhowmick S, D'Mello V, Caruso D, Abdul-Muneer PM Traumatic brain injury-induced downregulation of Nrf2 activates inflammatory response and apoptotic cell death J. Mol. Med. 2019-11-22 [PMID: 31758217]
• Lai A, Ho L, Evashwick-Rogler TW et al. Dietary polyphenols as a safe and novel intervention for modulating pain associated with intervertebral disc degeneration in an in-vivo rat model PLoS ONE 2019-10-02 [PMID: 31577822] (WB, Mouse)
• Singh WR, Devi HS, Kumawat S, et al. Angiogenic and MMPs modulatory effects of icariin improved cutaneous wound healing in rats Eur. J. Pharmacol. 2019-06-17 [PMID: 31220437] (WB, Rat)
• Song C, Wu YS, Yang ZY et al. Astrocyte-Conditioned Medium Protects Prefrontal Cortical Neurons from Glutamate-Induced Cell Death by Inhibiting TNF-a Expression Neuroimmunomodulation 2019-01-30 [PMID: 30699428] (WB, Rat)
• Fauzia E, Barbhuyan Tk, Shrivastava Ak et al. Chick Embryo: A Preclinical Model for Understanding Ischemia-Reperfusion Mechanism. Front Pharmacol. 2018-09-21 [PMID: 30298003] (Chicken)
• Evashwick-Rogler TW, Lai A, Watanabe H et al. Inhibiting tumor necrosis factor-alpha at time of induced intervertebral disc injury limits long-term pain and degeneration in a rat model. JOR Spine 2018-06-01 [PMID: 29963655] (Rat)
• Cesur MG, Gulle K, Sirin FB et al. Effects of curcumin and melatonin on bone formation in orthopedically expanded suture in rats: A biochemical, histological and immunohistochemical study Orthod Craniofac Res 2018-06-21 [PMID: 29927045] (IHC-P, Rat)
• Abdel-Zaher AO, Farghaly HSM, El-Refaiy AEM, Abd-Eldayem AM. Protective effect of the standardized leaf extract of Ginkgo biloba (EGb761) against hypertension-induced renal injury in rats Clin. Exp. Hypertens. 2018-01-19 [PMID: 29351002] (Rat)
• Al-Khalaf HH, Amir M, Al-Mohanna F et al. Obesity and p16INK4A Downregulation Activate Breast Adipocytes and Promote Their Protumorigenicity. Mol Cell Biol 2017-08-11 [PMID: 28630279] (Human)
• Gunel C, Demirci B, Eryilmaz A et al. Inhibitory Effect of Pycnogenol() on Airway Inflammation in Ovalbumin-Induced Allergic Rhinitis. Balkan Med J. 2016-11-01 [PMID: 27994914] (IF/IHC, Rat)
• Chen JY, Jian DY, Lien CC et al. Adipocytes play an etiological role in the podocytopathy of high-fat diet-fed rats. J. Endocrinol. 2016-08-18 [PMID: 27539963] (WB, Rat)
• Rossi AF, Cadamuro AC, Biselli-Perico JM et al. Interaction between inflammatory mediators and miRNAs in Helicobacter pylori infection. Cell. Microbiol. 2016-03-04 [PMID: 26945693] (IHC-P, Human)
• Wang X, Parrish AR. Loss of alpha(E)-catenin promotes Fas mediated apoptosis in tubular epithelial cells Apoptosis 2015-04-17 [PMID: 25894537] (WB, Rat)
• Tessier DR, Raha S, Holloway AC et al. Characterization of immune cells and cytokine localization in the rat utero-placental unit mid- to late gestation J. Reprod. Immunol. 2015-02-12 [PMID: 25725501] (IHC-P, Rat)
• Chen X, Zhang X, Wang Y et al. Inhibition of immunoproteasome reduces infarction volume and attenuates inflammatory reaction in a rat model of ischemic stroke Cell Death Dis. 2015-01-30 [PMID: 25633295] (WB, Rat)
• Rodriguez KA, Edrey YH, Osmulski P, Gaczynska M, Buffenstein R. Altered composition of liver proteasome assemblies contributes to enhanced proteasome activity in the exceptionally long-lived naked mole-rat. PLoS One;7(5):e35890. 2012-01-01 [PMID: 22567116] (WB, Mouse, Rat)
• Roth N, Stadler S, Lemann M, Hosli S, Simon HU, Simon D. Distinct eosinophil cytokine expression patterns in skin diseases - the possible existence of functionally different eosinophil subpopulations. Allergy;66(11):1477-86. 2011-11-01 [PMID: 21884530] (IF/IHC, Human)

Review for TNF-alpha Antibody (NB600-587) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Rat.

reviewed by: Verified Customer WB Rat 05/22/2015

Summary

• Application: Western Blot
• Sample Tested: NRK whole cell lysate
• Species: Rat

FAQs for TNF-alpha Antibody (NB600-587). (Showing 1 - 4 of 4 FAQs).

1. I need to know about TNF-alpha antibody kits for mice.
   • We currently have two TNFalpha ELISA kits specific for mouse: catalog numbers KA0257 and NBP1-92670.
2. I would like to ask you for help. I need an antibody for Elisa to detect human TNF alpha in a supernatant from a cell culture (meaning supernatant after centrifugation of collected cell suspension from a plate well). Which of your antibodies against human tnf alpha would be suitable? I would like to buy only the primary antibody.
   • I would recommend catalog number NBP1-67821 or NB600-587; however, a full list of our anti-human TNF alpha antibodies suitable for use in ELISA can be found using this link.
3. I am interested in a TNF alpha antibody, cross reactive for human, rat and mouse (host species: rabbit). Could your product NBP1-19532 be used in western blotting applications? Or do you have a similar product in your catalog which could fit with my request?
   • The antibody you mention, NBP1-19532, has not yet been validated in Western blot. I would instead recommend either NBP1-67821 or NB600-587. These are both rabbit polyclonal antibodies that cross-reacts with human, mouse and rat and have been used in Western blotting.
4. Can your TNF-alpha products be used to treat TBI victims and therefore avoid the perispinal injections with Enbrel?
   • I am very sorry, but all of our products are for scientific research use only, and none are intended or approved for use in humans.

Bioinformatics

• Gene Symbol: TNF
• Entrez:
  • 7124(Human)
• Uniprot:
  • P01375()
  • P01375()
  • P01375()
  • P01375()