Human TNF-alpha ELISA Kit (Colorimetric)

Novus Biologicals | Catalog # NBP1-91170

Novus Biologicals
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Key Product Details

Sample Type & Volume Required Per Well

Cell culture supernatant, serum, plasma (EDTA, citrate, heparin), urine (50 uL)

Sensitivity

2.3 pg/mL

Assay Range

7.8 - 500 pg/mL
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Product Specifications

Assay Type

Sandwich ELISA

Kit Type

ELISA Kit (Colorimetric)

Species

Human

Specificity

The interference of circulating factors of the immune system was evaluated by spiking these proteins at physiologically relevant concentrations into a human TNF alpha positive serum. There was no interference detected, namely not with TNF-R (60 kDa and 80 kDa).

Description

The human TNF-alpha ELISA is an enzyme-linked immunosorbent assay for the quantitative detection of human TNF-alpha

Precision

Intra-Assay Precision (Precision within an assay) CV% 6.0%

Inter-Assay Precision (Precision between assays) CV% 7.4%

Recovery for Human TNF-alpha ELISA Kit (Colorimetric)

Recovery

The recovery ranged from 72% to 112% with an overall mean recovery of 93%. Please see protocol for more specific Spike Recovery details.

Scientific Data Images for Human TNF-alpha ELISA Kit (Colorimetric)

ELISA: Human TNF-alpha ELISA Kit (Colorimetric) [NBP1-91170]

ELISA: Human TNF-alpha ELISA Kit (Colorimetric) [NBP1-91170]

ELISA: TNF-alpha ELISA Kit [NBP1-91170] - Standard curve

Kit Contents for Human TNF-alpha ELISA Kit (Colorimetric)

  • Adhesive Films
  • Aluminium pouch(es) with a Microwell Plate coated with monoclonal antibody to human TNF-alpha
  • Assay Buffer Concentrate 20x (PBS with 1% Tween 20 and 10% BSA)
  • Biotin-Conjugate anti-human TNF-alpha polyclonal antibody
  • Control high, lyophilized
  • Control low, lyophilized
  • Human TNF-alpha Standard lyophilized, 1000 pg/mL upon reconstitution
  • Sample Diluent
  • Stop Solution (1M Phosphoric acid)
  • Streptavidin-HRP
  • Substrate Solution (tetramethyl-benzidine)
  • Wash Buffer Concentrate 20x (PBS with 1% Tween 20)

Preparation and Storage

Shipping

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

Stability & Storage

Storage of components varies. See protocol for specific instructions.

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 Human TNF-alpha ELISA Kit (Colorimetric)

Certificate of Analysis

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

Product Specific Notices for Human TNF-alpha ELISA Kit (Colorimetric)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. ELISA Kits are guaranteed for 6 months from date of receipt.

Citations for Human TNF-alpha ELISA Kit (Colorimetric)

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FAQs for Human TNF-alpha ELISA Kit (Colorimetric)

Showing  1 - 4 of 4 FAQs Showing All

    • 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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