Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Canine, Fish

Cited:

Human, Mouse, Rat, Canine, Fish, Primate - Macaca mulatta (Rhesus Macaque)

Predicted:

Baboon (97%), Feline (90%), Monkey (98%), Primate (100%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Immunohistochemistry Free-Floating, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Immunohistochemistry Free-Floating, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all TNF-alpha Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to an internal portion of the human TNF alpha protein (between residues 100-200) [Uniprot: P01375]

Reactivity Notes

Fish reactivity reported in scientific literature (PMID: 29321977). Immunogen displays the following percentage of sequence identity for non-tested species: porcine (89%), equine (87%), and guinea pig (84%). Use in Canine reported in scientific literature (PMID:32656339).

Localization

Membrane

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

Novus Biologicals Rabbit TNF-alpha Antibody - BSA Free (NBP1-19532) is a polyclonal antibody validated for use in IHC, WB, Flow and ICC/IF. Anti-TNF-alpha Antibody: Cited in 82 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for TNF-alpha Antibody - BSA Free

Western Blot: TNF-alpha AntibodyBSA Free [NBP1-19532]

Western Blot: TNF-alpha AntibodyBSA Free [NBP1-19532]

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 AntibodyBSA Free [NBP1-19532]

Western Blot: TNF-alpha AntibodyBSA Free [NBP1-19532]

TNF-alpha-Antibody-Western-Blot-NBP1-19532-img0008.jpg
Immunohistochemistry-Paraffin: TNF-alpha Antibody - BSA Free [NBP1-19532]

Immunohistochemistry-Paraffin: TNF-alpha Antibody - BSA Free [NBP1-19532]

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 - BSA Free [NBP1-19532]

Immunohistochemistry-Paraffin: TNF-alpha Antibody - BSA Free [NBP1-19532]

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.
TNF-alpha Antibody - BSA Free

Western Blot: TNF-alpha Antibody - BSA Free [NBP1-19532] -

Scutellarin attenuates hypertension-induced brain expression of NF-kappa B, TNF-alpha, IL-1 beta, and IL-18. Western immunoblot analysis for (a) NF-kappa B p65, (b) TNF-alpha, (c) IL-1 beta, and (d) IL-18 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.
TNF-alpha Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: TNF-alpha Antibody - BSA Free [NBP1-19532] -

Immunocytochemistry/ Immunofluorescence: TNF-alpha Antibody - BSA Free [NBP1-19532] - Effects of femoral artery occlusion on TNF-alpha signal in sensory nerves. (A) Seventy-two hours of femoral artery occlusion increased the levels of TNF-alpha in the DRG tissues as compared with them in DRG of control limbs (n = 6 in each group); & PTX given into the hindlimb muscles attenuated amplification of TNF-alpha in the DRG tissues of limbs with femoral artery occlusion (n = 8). ∗P < 0.05, occlusion group vs. control & occlusion with prior PTX. (B) Immunofluorescence was used to examine double-labeling for TNF-alpha & peripherin/NF-200 (n = 3). Peripherin was used to label DRG neurons that project thin C-fibers. NF200 was used to identify A-fibers of DRG neurons. Representative photomicrographs show co-existence of TNF-alpha & peripherin staining in DRG neurons (top panel), whereas few TNF-alpha & NF-200 staining were observed in DRG neurons (bottom panel). Arrows indicate representative positive cells for both TNF-alpha & peripherin after they were merged. Scale bar = 50 μm. (C) Representative bands (left panel) & averaged data (right panel), demonstrating that femoral artery occlusion upregulated protein expression TNF-alpha receptor subtype TNFR1, but not TNFR2. A significant difference in TNFR1 was seen between control & occluded groups. ∗P < 0.05 vs. control. n = 6 in each group. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30374312), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TNF-alpha Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: TNF-alpha Antibody - BSA Free [NBP1-19532] -

Immunocytochemistry/ Immunofluorescence: TNF-alpha Antibody - BSA Free [NBP1-19532] - Immunoflorescence staining results of TNF-alpha (96 h). A). Lamella of gill tissue (arrow). Control group. IF. 20 μm. B) Positive reaction in lamellar cells of gill tissue (arrows). Low dose toxic group. IF. 20 μm. C) Positivity in lamellar cells of gill tissue (arrow). High dose toxic group. IF. 20 μm. D) Liver tissue, vena centralis (arrow). Control group. IF. 20 μm. E) Positive reactions in hepatocytes (arrows). Low dose toxic group. IF. 20 μm. F) Positive reactions in hepatocytes (arrows). High dose toxic group. IF. 20 μm. G) Weak positive reactions in neurons (arrows). Control group. IF. 20 μm. H) Weak positivity in neurons (arrows). Low dose toxic group. IF. 20 μm. I) Positivity in neurons (arrows). High dose toxic group. IF. 20 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29321977), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for TNF-alpha Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:200 - 1:300

Immunohistochemistry

1:200

Immunohistochemistry-Paraffin

1:200

Western Blot

1:250 - 1:500
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).

Reviewed Applications

Read 3 reviews rated 5 using NBP1-19532 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: TNF-alpha

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

Long Name

Tumor Necrosis Factor alpha

Alternate Names

Cachetin, DIF, TNF, TNF-A, TNFA, TNFalpha, TNFG1F, TNFSF1A, TNFSF2

Entrez Gene IDs

7124 (Human); 21926 (Mouse); 24835 (Rat)

Gene Symbol

TNF

UniProt

P01375

Additional TNF-alpha Products

Product Documents for TNF-alpha Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for TNF-alpha Antibody - BSA Free

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.

Citations for TNF-alpha Antibody - BSA Free

Customer Reviews for TNF-alpha Antibody - BSA Free (3)

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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • TNF-alpha Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Primary neurons
    Species: Rat
    Verified Customer | Posted 10/13/2016
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: trabecular meshwork (TM) region of pig eyes
    Species: Other
    Verified Customer | Posted 01/07/2016
    Pig TM tissue stained with TNF alpha antibody
    TNF-alpha Antibody - BSA Free NBP1-19532
  • Name: Ranveer Singh
    Application: Western Blot
    Sample Tested: Recombinant his-tagged TNFa made in E. coli cells
    Species: Human
    Verified Customer | Posted 07/24/2014
    WB for His-tagged TNFa protein made in E. coli
    TNF-alpha Antibody - BSA Free NBP1-19532

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Protocols

View specific protocols for TNF-alpha Antibody - BSA Free (NBP1-19532):


Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.


Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for TNF-alpha Antibody - BSA Free

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    • Q: Can your TNF-alpha products be used to treat TBI victims and therefore avoid the perispinal injections with Enbrel?
      A: 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.

    • Q: 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?
      A: 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.

    • A: We currently have two TNFalpha ELISA kits specific for mouse: catalog numbers KA0257 and NBP1-92670.

    • Q: 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.
      A: 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.

    • Q: I would like to learn about the TNF alpha antibody (NBP1-19532) immunohistochemistry protocol. Is there any specific protocol you have?
      A: Our lab did not use any customized protocol for the IHC-P validation of this antibody. Please see our general IHC-P protocol.

    • Q: Is there any positive control tissue besides human colon carcinoma tissue that can be used for for paraffin embedded IHC testing?
      A: You can use skin, lung, liver or mammary tumor tissue etc as a positive control for your assay.

    • Q: 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.
      A: 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.
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