TNF-alpha Antibody - Azide and BSA Free

Western Blot: TNF-alpha Antibody [NBP2-44062] - Membrane blocked in 1% BSA-TBS-T 30 min RT, Rb-a-TNF alpha added at 1:1000 in 1% BSA-TBS-T o/n 4C, DyLight 649 Goat anti-Rabbit added at 1:20,000 in blocking buffer for 30 min RT.

Immunohistochemistry: TNF-alpha Antibody [NBP2-44062] - Human prostate tissue was stained using TNFa antibody. This image was submitted via customer Review.

Immunohistochemistry: TNF-alpha Antibody [NBP2-44062] - Fluorescent immunohistochemistry showing staining of human colon.

Immunohistochemistry: TNF-alpha Antibody [NBP2-44062] - 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.

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 antibody. Marker: Opal Pre-stained ladder

Western blot using Anti-Human TNF-a (RABBIT) Antibody. Membrane blocked in 1% or 30 min at RT, Rb-a-TNF alpha added at 1:1000 in 1% /n 4C, DyLight 649 Gt-a-Rb

• Reactivity: Hu
• Applications: WB, ELISA, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: Azide and BSA Free

TNF-alpha Antibody - Azide and BSA Free Summary

• Description: Store this antibody 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: TNF-alpha Antibody used to produce this IgG fraction antibody was prepared by repeated immunizations with recombinant human TNFa. (Uniprot: P01375)
• Specificity: This antibody is primarily directed against mature 17,000 MW human TNFalpha and is useful in determining its presence in various assays. In general, this antibody also detects primate TNFalpha in the same formats using similar dilutions. The antibody does not recognize human TNFbeta (lymphotoxin). This IgG fraction antibody will recognize the cell-bound precursor of TNFalpha as a 26,000 protein in immunoblots, particularly in denatured samples. This antibody is also useful for neutralization of human and primate TNFalpha 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: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: TNF
• Purity: IgG purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:1000-1:5000
  • Immunocytochemistry/ Immunofluorescence 1:10 - 1:500
  • Immunohistochemistry 1:100-1:500
  • Immunohistochemistry-Paraffin
  • Western Blot 1:500-1:2000
• Application Notes: This product has been tested in WB, ELISA, IF, and IHC. Expect a band in appropriate lysates at ~25.6kDa. This antibody is also useful for neutralization of human and primate TNFalpha activity in bioassays diluted at 1:200. The antibody does not recognize human TNF beta (lymphotoxin). Specific conditions for reactivity and signal detection should be optimized by the end user.
  
  IHC reported in a verified customer review.

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Preservative: No Preservative
• Purity: IgG purified

Alternate Names for TNF-alpha Antibody - Azide and BSA Free

• 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.

Review for TNF-alpha Antibody (NBP2-44062) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Sachin Puri IHC Human 06/15/2017

Summary

• Application: Immunohistochemistry
• Sample Tested: Prostate tissue
• Species: Human
• Lot: N/A

FAQs for TNF-alpha Antibody (NBP2-44062). (Showing 1 - 4 of 4 FAQ).

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
• Uniprot:
  • P01375()
  • P01375()
  • P01375()
  • P01375()