Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Mouse

Applications

Block/Neutralize, CyTOF-ready, Flow Cytometry, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 6N1E7

Format

Azide Free

Concentration

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

View all TNF-alpha Antibodies »

Product Specifications

Immunogen

Full-length recombinant human TNF alpha was used as the immunogen for this antibody.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Scientific Data Images for TNF-alpha Antibody (6N1E7) - Azide Free

Western Blot: TNF-alpha Antibody (6N1E7)Azide Free [NBP2-27224]

Western Blot: TNF-alpha Antibody (6N1E7)Azide Free [NBP2-27224]

Western Blot: TNF-alpha Antibody (6N1E7) - Azide Free [NBP2-27224] - Analysis of TNF alpha recombinant protein using TNF alpha antibody at 3 ug/ml. Goat anti-mouse Ig HRP secondary antibody and PicoTect ECL substrate solution were used for this test.
Flow Cytometry: TNF-alpha Antibody (6N1E7) - Azide Free [NBP2-27224]

Flow Cytometry: TNF-alpha Antibody (6N1E7) - Azide Free [NBP2-27224]

Flow Cytometry: TNF-alpha Antibody (6N1E7) - Azide Free [NBP2-27224] - Human PBMC were stimulated with PMA (10 ng/ml) and Ionomycin (500 ng/ml) for 5h in presence of Brefeldin A. At the end of stimulation, cells were surface stained with before testing with by intracellular staining. Fixed and permeabilized cells were incubated with 10 ul (0.5 ug) of TNF antibody (or ) for 30 minutes on ice. Lymphocyte population was gated for analysis to show TNF alpha positive T cells.
Blocking/Neutralizing: TNF-alpha Antibody (6N1E7) - Azide Free [NBP2-27224] - Validation of inhibitory effects of TNF alpha antibody on TNF alpha activity in the cell line. NF-kB/SEAPorter cells were plated in 96-well plate at 5 x 10^4 cells per well for 16 h. Cells were pre-incubated with TNF alpha antibody at 10 and 20 ug/ml for 1 hr, then treated with 10 ng/ml TNF alpha or 10 ng/ml PMA. After 24h, the secreted alkaline phosphatase (SEAP) was analyzed using the SEAPorterAssay Kit .

Applications for TNF-alpha Antibody (6N1E7) - Azide Free

Application
Recommended Usage

Flow Cytometry

1ul/1 million cells

Western Blot

2-4ug/ml
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

0.2 ml sterile PBS

Format

Azide Free

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 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

Gene Symbol

TNF

Additional TNF-alpha Products

Product Documents for TNF-alpha Antibody (6N1E7) - Azide Free

Product Datasheets

Certificate of Analysis

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

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Product Specific Notices for TNF-alpha Antibody (6N1E7) - Azide 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.

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