TNF-alpha Antibody

Novus Biologicals | Catalog # NB600-587

Novus Biologicals
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Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse, Rat, Chicken, Primate

Cited:

Human, Mouse, Rat, Avian - Chicken

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit Serum
View all TNF-alpha Primary Antibodies »

Product Specifications

Immunogen

The whole rabbit serum was prepared by repeated immunizations with recombinant human TNF-alpha produced in E.coli. (Uniprot: P01375)

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).

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.

Clonality

Polyclonal

Host

Rabbit

Isotype

Serum

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.

Scientific Data Images for TNF-alpha Antibody

Western Blot: TNF-alpha AntibodyAzide Free [NB600-587]

Western Blot: TNF-alpha AntibodyAzide Free [NB600-587]

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 AntibodyAzide Free [NB600-587]

Western Blot: TNF-alpha AntibodyAzide Free [NB600-587]

TNF-alpha-Antibody---Azide-Free-Western-Blot-NB600-587-img0014.jpg
Immunocytochemistry/ Immunofluorescence: TNF-alpha Antibody - Azide Free [NB600-587]

Immunocytochemistry/ Immunofluorescence: TNF-alpha Antibody - Azide Free [NB600-587]

TNF-alpha-Antibody---Azide-Free-Immunocytochemistry-Immunofluorescence-NB600-587-img0015.jpg
Immunohistochemistry: TNF-alpha Antibody - Azide Free [NB600-587]

Immunohistochemistry: TNF-alpha Antibody - Azide Free [NB600-587]

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 AntibodyAzide Free [NB600-587]

Western Blot: TNF-alpha AntibodyAzide Free [NB600-587]

TNF-alpha-Antibody---Azide-Free-Western-Blot-NB600-587-img0013.jpg
TNF-alpha Antibody - Azide Free

TNF-alpha Antibody - Azide Free

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.
TNF-alpha Antibody - Azide Free

TNF-alpha Antibody - Azide Free

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.
TNF-alpha Antibody - Azide Free

TNF-alpha Antibody - Azide Free

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

Applications for TNF-alpha Antibody

Application
Recommended Usage

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.

Reviewed Applications

Read 1 review rated 5 using NB600-587 in the following applications:

Formulation, Preparation, and Storage

Purification

Whole Antiserum

Formulation

Antiserum

Preservative

No Preservative

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

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

Stability & Storage

Store at -20C short term. Aliquot and store at -80C 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)

Gene Symbol

TNF

UniProt

P01375

Additional TNF-alpha Products

Product Documents for TNF-alpha Antibody

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

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

Customer Reviews for TNF-alpha Antibody (1)

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  • Name: Anonymous
    Application: Western Blot
    Sample Tested: NRK whole cell lysate
    Species: Rat
    Verified Customer | Posted 05/22/2015
    Western Blot of TNF-alpha in NRK cells
    TNF-alpha Antibody NB600-587

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Protocols

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FAQs for TNF-alpha Antibody

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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.
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