Rat TNF-alpha ELISA Kit (Colorimetric)

Novus Biologicals | Catalog # NBP1-92681

Novus Biologicals
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Key Product Details

Sample Type & Volume Required Per Well

Cell culture supernatant, serum (50 uL)

Sensitivity

11.0 pg/mL

Assay Range

39.1 - 2,500 pg/mL
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Product Specifications

Assay Type

Sandwich ELISA

Kit Type

ELISA Kit (Colorimetric)

Species

Rat

Specificity

The interference of circulating factors of the immune systemwas evaluated by spiking these proteins at physiologically relevant concentrations into a rat TNF alpha positive serum. There was no crossreactivity detected.

Description

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

Precision

Intra-Assay Precision (Precision within an assay) CV% 5.0%

Inter-Assay Precision (Precision between assays) CV% 10%

Recovery for Rat TNF-alpha ELISA Kit (Colorimetric)

Recovery

109%

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

ELISA: Rat TNF-alpha ELISA Kit (Colorimetric) [NBP1-92681]

ELISA: Rat TNF-alpha ELISA Kit (Colorimetric) [NBP1-92681]

ELISA: TNF-alpha ELISA Kit [Biotin] [NBP1-92681] - Standard curve

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

  • Adhesive Films
  • Aluminium pouch(es) with a Microwell Plate coated with polyclonal antibody to rat TNF-alpha
  • Assay Buffer Concentrate 20x (PBS with 1% Tween 20 and 10% BSA)
  • Biotin-Conjugate anti-rat TNF-alpha polyclonal antibody
  • Rat TNF-alpha Standard lyophilized, 5 ng/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 Rat TNF-alpha ELISA Kit (Colorimetric)

Certificate of Analysis

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

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FAQs for Rat TNF-alpha ELISA Kit (Colorimetric)

Showing  1 - 5 of 5 FAQs Showing All

    • Q: Can we use NBP1-92681 (Rat TNF-alpha ELISA Kit, Biotin) on rat tissue lysate? If yes, please share how to prepare the lysate?
      A: Yes, this kit is for rat samples especially the cell culture supernatant and serum but it should be good for rat tissue lysate also. We have not tested it on tissue lysates yet, but we have had reports from other customers using lysis buffers containing 0.5% TX-100 showing no deleterious effect on the performance of some of our kits. We do recommend that as a positive control you spike in some of the standard at a known concentration in your buffer and see if it performs as expected. In short, the lab recommended to use one or more cell line supernatants as the positive control in the ELISA assays.

    • 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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