TNF-alpha Antibody

Western Blot: TNF-alpha Antibody [NBP1-19532] - Recombinant human TNF alpha (10 ng) was separated on a 12% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-TNF alpha in 5% block buffer and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.

Western Blot: TNF-alpha Antibody [NBP1-19532] - Scutellarin attenuates hypertension-induced brain expression of NF-kappaB, TNF-alpha, IL-1beta, and IL-18. Western immunoblot analysis for TNF-alpha in the rat cortex and striatum. Treatment with scutellarin significantly reduced the expression of these inflammatory markers in a dose-dependent manner. *P < 0.001 versus sham group; #P < 0.05, P < 0.001 versus NS group; &P < 0.001 versus low-dose group. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/mi/2013/432623/), licensed under a CC-BY licence.

Immunocytochemistry/Immunofluorescence: TNF-alpha Antibody [NBP1-19532] - TNF alpha antibody was tested in A431 cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).

Immunohistochemistry-Paraffin: TNF-alpha Antibody [NBP1-19532] - Analysis of a FFPE tissue section of mouse intestine using TNF-alpha antibody (NBP1-19532) at 1:300 dilution. The binding of this primary antibody to TNF-alpha protein in the section was detected using HRP-labeled secondary antibody and DAB reagent, and nuclei of cells were counterstained using hematoxylin. This TNF-alpha antibody generated an expected diffused immunostaining of this protein in the tested tissue. Staining was primarily observed in the epithelial cells and some cells showed membrane positivity also.

Immunohistochemistry-Paraffin: TNF-alpha Antibody [NBP1-19532] - Analysis of a FFPE tissue section of mouse intestine using TNF-alpha antibody (NBP1-19532) at 1:300 dilution. The binding of this primary antibody to TNF-alpha protein in the section was detected using HRP-labeled secondary antibody and DAB reagent, and nuclei of cells were counterstained using hematoxylin. This TNF-alpha antibody generated an expected immunopositivity of this protein in the tested tissue. Staining was primarily observed in the epithelial cells while some staining was present in mucosa muscularis also.

• Reactivity: Hu, Mu, Rt, Fi, Pm, Ca, Fe, Pm, Pm
• Applications: WB, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IHC-FrFl
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

TNF-alpha Antibody Summary

• Immunogen: A synthetic peptide made to an internal portion of the human TNF alpha protein (between residues 100-200) [Uniprot: P01375]
• Localization: Membrane
• Predicted Species: Primate (100%), Monkey (98%), Canine (94%), Feline (90%), Baboon (97%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: TNF
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:250 - 1:500
  • Flow Cytometry
  • Immunocytochemistry/Immunofluorescence 1:200 - 1:300
  • Immunohistochemistry 1:200
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:200
  • Immunohistochemistry Free-Floating
• Application Notes: Western Blot data from customer review and citation (PMID: 24223475). Use in Immunohistochemistry-free floating reported in scientific literature (PMID: 26219646). Use in Immunohistochemistry-Frozen reported in scientific literature (PMID: 26725948). Use in FLOW reported in scientific literature (PMID: 26889045).

Reactivity Notes

Fish reactivity reported in scientific literature (PMID: 29321977). Immunogen has 89% homology with porcine, 87% with equine, and 84% with guinea pig.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

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 Alpha (TNF alpha) is a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. This cytokine is mainly secreted by macrophages and causes tumor necrosis when injected into tumour bearing mice. TNF alpha exists as a multimer of two, three, or five noncovalently linked units, but shows a single 17 kDa band following SDS PAGE under non reducing conditions. TNF alpha is closely related to the 25 kDa protein Tumour Necrosis Factor beta (lymphotoxin), sharing the same receptors and cellular actions. It can bind to, and thus functions through its receptors TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. This cytokine is involved in the regulation of a wide spectrum of biological processes including cell proliferation, differentiation, apoptosis, lipid etabolism, and coagulation and it has been implicated in a variety of diseases, including autoimmune diseases, insulin resistance, and cancer. Knockout studies in mice also suggested the europrotective function of this cytokine. TNF alpha is currently being evaluated in treatment of certain cancers and AIDS Related Complex.

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 (NBP1-19532)(28)

Publications using NBP1-19532

• Han Y, Lee Y, Jang Y et Al. Aspirin Improves Nonalcoholic Fatty Liver Disease and Atherosclerosis through Regulation of the PPARdelta-AMPK-PGC-1 alpha Pathway in Dyslipidemic Conditions BioMed Research International Mar 19 2020 [PMID: 32258142] (WB, Human, Mouse)
• Wang M, Zong Hf, Chang Kw et Al. 5-HT1AR alleviates A beta-induced cognitive decline and neuroinflammation through crosstalk with NF-kappa B pathway in mice Int. Immunopharmacol. Jan 1 2020 [PMID: 32143008] (IHC, Mouse)
• Fu XQ, Peng J, Wang AH, Luo ZG Tumor necrosis factor alpha mediates neuromuscular synapse elimination Cell Discov Mar 3 2020 [PMID: 32140252] (IHC, Mouse)
• Wang M, Law ME, Davis BJ, et al. Identification of hub genes and key pathways of dietary advanced glycation end products induced non alcoholic fatty liver disease by bioinformatics analysis and animal experiments Mol Med Report Dec 9 2019 [PMID: 31974594] (IHC, Mouse)
• Li X, Xue YM, Guo HM et al. High hydrostatic pressure induces atrial electrical remodeling through upregulation of inflammatory cytokines Life Sci. Dec 20 2019 [PMID: 31870776] (WB, Mouse, Rat)
• Sparks RP, Lui A, Bader D et al. A specific small-molecule inhibitor of Protein Kinase CdeltaI activity improves metabolic dysfunction in human adipocytes from obese individuals J. Biol. Chem. Aug 14 2019 [PMID: 31413114] (WB, Human)
• Li S, Jiang L, Yang Y et al. Siglec1 enhances inflammation through miR-1260-dependent degradation of I kappa B alpha in COPD Exp. Mol. Pathol. Jan 30 2020 [PMID: 32007531] (WB, Rat)
• Corbett BF, Luz S, Arner J et al. Sphingosine-1-phosphate receptor 3 in the medial prefrontal cortex promotes stress resilience by reducing inflammatory processes Nat Commun Jul 17 2019 [PMID: 31316053] (IHC-Fr, Rat)
• Gu X, Peng CY, Lin SY et al. P16INK4a played a critical role in exacerbating acute tubular necrosis in acute kidney injury Am J Transl Res Jun 15 2019 [PMID: 31312394] (WB, Mouse)
• Bicakci, N;Karaboga, I;Dokmeci, AH;Guzel, S;Fidanol Erboga, Z; Cardioprotective effect of caffeic acid phenethyl ester on cardiac contusion following blunt chest trauma in rats Biotech Histochem Apr 1 2019 [PMID: 30929510] (IHC, Rat)
• Xing J, Lu J, Li J. Role of TNF-a in Regulating the Exercise Pressor Reflex in Rats With Femoral Artery Occlusion. Front Physiol. Oct 15 2018 [PMID: 30374312] (ICC/IF, Rat)
• Park SH, Rhee J, Kim SK et al. BATF regulates collagen-induced arthritis by regulating T helper cell differentiation Arthritis Res. Ther. Aug 2 2018 [PMID: 30071881] (ICC/IF, IHC-P, Mouse)
• Hoshikawa H, Kamitori K, Indo K, Mori T. Combined treatment with D-allose, docetaxel and radiation inhibits the tumor growth in anin vivomodel of head and neck cancer. Oncol Lett. 2018 Mar 01 [PMID: 29456721] (IHC, Human)
• Ozdemira S, Altunb S, Arslan H. Imidacloprid exposure cause the histopathological changes, activation of TNF-a, iNOS, 8-OHdG biomarkers, and alteration of caspase 3, iNOS, CYP1A, MT1 gene expression levels in common carp (Cyprinus carpio L.). Toxicology Reports 2017 [PMID: 29321977] (ICC/IF, Fish)
• Mohamed R, Coucha M, Elshaer SL et al. Inducible overexpression of endothelial proNGF as a mouse model to study microvascular dysfunction. Biochim. Biophys. Acta 2017 Dec 16 [PMID: 29253516] (Mouse)
• Huegel J, Choi DS, Nuss CA et al. Effects of pulsed electromagnetic field therapy at different frequencies and durations on rotator cuff tendon-to-bone healing in a rat model J Shoulder Elbow Surg 2017 Nov 22 [PMID: 29174271] (IHC, Rat)
• Veruva SY, Lanman TH, Isaza JE et al. Periprosthetic UHMWPE Wear Debris Induces Inflammation, Vascularization, and Innervation After Total Disc Replacement in the Lumbar Spine. Clin. Orthop. Relat. Res. Aug 3 2016 [PMID: 27488379] (IHC, Mouse)
• Szylberg, Janiczek M, Popiel A, Marszatek A et al. Expression of COX-2, IL-1beta, TNF-alpha and IL-4 in epithelium of serrated adenoma, adenoma and hyperplastic polyp Arch Med Sci Feb 1 2016 [PMID: 26925134] (IHC-P, Human)
• Xu Y, Romero R, Miller D et al. An M1-like Macrophage Polarization in Decidual Tissue during Spontaneous Preterm Labor That Is Attenuated by Rosiglitazone Treatment J. Immunol. Feb 17 2016 [PMID: 26889045] (IHC-Fr, FLOW, Human)
• Vieira Ramos G, Pinheiro CM, Messa SP et al. Cryotherapy Reduces Inflammatory Response Without Altering Muscle Regeneration Process and Extracellular Matrix Remodeling of Rat Muscle. Sci Rep. 2016 Jan 04 [PMID: 26725948] (IHC-Fr, Rat)
  
  Details:
  TNF alpha antibody (NBP1-19532) was used at 1:200 dilution for IHC-Fr staining of Tibialis anterior/TA muscle of rats 3, 7 and 14 days post-injury (subjected to freezing muscle injury and cryotherapy). TNF alpha expression was observed in CD68+ve cells and in injured muscle fibers.
• Acosta SA, Tajiri N, Hoover J et al. Intravenous Bone Marrow Stem Cell Grafts Preferentially Migrate to Spleen and Abrogate Chronic Inflammation in Stroke. Stroke. 2015 Sep 01 [PMID: 26219646] (IHC-FrFl, Rat)
• Tucker JJ, Riggin CN, Connizzo BK et al. Effect of overuse-induced tendinopathy on tendon healing in a rat supraspinatus repair model J. Orthop. Res. 2015 Jul 27 [PMID: 26218457] (IHC-P, Rat)
• Castoldi G, di Gioia CR, Bombardi C et al. PREVENTION OF DIABETIC NEPHROPATHY BY COMPOUND 21, SELECTIVE AGONIST OF THE ANGIOTENSIN TYPE 2 RECEPTORS, IN ZUCKER DIABETIC FATTY RATS. Am. J Physiol. Renal Physiol. 2014 Sep 03 [PMID: 25186297]
• Thomas SJ, Sarver JJ, Yannascoli SM et al. Effect of isolated hyperglycemia on native mechanical and biologic shoulder joint properties in a rat model. J. Orthop. Res. 2014 Jul 28 [PMID: 25070655] (IHC-P, Rat)
  
  Details:
  TNF alpha/TNFa antibody used for IHC-P on whole shoulders of adult male Sprague-Dawley rats subjected to STZ induced hyperglycemia for 8 weeks - shoulders fixed in formalin, cut 7um longitudinal sections, peroxidase blocked in 3% H2O2/methanol 48C for 30 min, non-specific protein blocked with 4% dried milk in PBS /Blotto for 20 min, primary used at 1:750 dilution/PBS and incubated 4C ON followed by detection via goat anti-rabbit peroxidase-conjugated/HRP - DAB method. Immunostaining image not shown, but staining score on Supraspinatus tendon and Superior capsule tissues shown in Table 2.
• Piao HL, Yuan Y, Wang M et al. alpha-catenin acts as a tumour suppressor in E-cadherin-negative basal-like breast cancer by inhibiting NF-kB signalling. Nat. Cell Biol. 2014 Feb 28 [PMID: 24509793] (IHC-P, Human)
• Chen X, Shi X, Zhang X et al. Scutellarin attenuates hypertension-induced expression of brain toll-like receptor 4/nuclear factor kappa B. Mediators Inflamm. 2013 [PMID: 24223475] (WB, Rat)
• Kwon KY, Jung HR, Hwang I, Choi WI. Evaluation of Bronchiolar and Alveolar Cell Injuries Induced by Short- and Long-term Exposure to Sidestream Smoke. Korean J Pathol 2012 Apr [PMID: 23109995]

Reviews for TNF-alpha Antibody (NBP1-19532) (3)

Average Rating: 5

(Based on 3 reviews)

We have 3 reviews tested in 3 species: Human, Rat, Other.

reviewed by: Anonymous ICC/IF Rat 10/13/2016

Summary

• Application: Immunocytochemistry/Immunofluorescence
• Sample Tested: Primary neurons
• Species: Rat

reviewed by: Anonymous IF Other 01/07/2016

Summary

• Application: Immunofluorescence
• Sample Tested: trabecular meshwork (TM) region of pig eyes
• Species: Other

Blocking

• Blocking Details: -

Primary Anitbody

• Dilution Ratio: 2 ug/ml

Secondary Antibody

• Secondary Description: Alexa 594

Details

• Detection Notes: -
• Fixation Details: 2% paraformaldehyde fixed for 20 minutes

Comments

• Comments: We used fresh pig eyes, then cut and keep trabecular meshwork (TM) region and 2 percent paraformaldehyde fixed for 20 minutes, then standard IHC was carried out.

reviewed by: Ranveer Singh WB Human 07/24/2014

Summary

• Application: Western Blot
• Sample Tested: Recombinant his-tagged TNFa made in E. coli cells
• Species: Human

Blocking

• Blocking Details: 5% BSA in TBST for 1 h

Primary Anitbody

• Dilution Ratio: 1:2000, Overnight at 4 degree in TBST on a rocker

Secondary Antibody

• Secondary Description: Goat anti-rabbit IgG HRP (Biorad)
• Secondary Manufacturer Cat#: 170-5046
• Secondary Concentration: 1:20,000 in 5% BSA

Details

• Detection Notes: ECL reagent was used. Exposure for 5 sec

FAQs for TNF-alpha Antibody (NBP1-19532). (Showing 1 - 7 of 7 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.
5. We ordered a TNF alpha antibody from you back in May, and I am inquiring about the expiration date. The antibody we ordered was cat #NBP1-19532 and lot # 360986.. The data sheet says that short term storage at 4C and -20C for long term storage but that they are only guaranteed for 6 months. My question is are you considering the "short term storage" at 4C to be the 6 month expiration and -20C the antibody should still be OK? Typically we give our antibodies that are stored at -20C a two year expiration if one is not given.
   • We suggest to store the antibody at 4 degrees for no longer than a week. After this the product should be stored at -20 degrees and we are not sure if this is can be used after the 6 month period. It is likely that the antibody should be fine however we only guarantee our products for up to 6 months.
6. Is there any positive control tissue besides human colon carcinoma tissue that can be used for for paraffin embedded IHC testing?
   • You can use skin, lung, liver or mammary tumor tissue etc as a positive control for your assay.
7. I would like to learn about the TNF alpha antibody (NBP1-19532) immunohistochemistry protocol. Is there any specific protocol you have?
   • Our lab did not use any customized protocol for the IHC-P validation of this antibody. Please see our general IHC-P protocol.

Bioinformatics

• Gene Symbol: TNF
• Entrez:
  • 7124(Human)
  • 21926(Mouse)
  • 24835(Rat)
• Uniprot:
  • P01375()