Human TNF alpha ELISA

The TNF alpha ELISA is an immunoassay kit for quantitative measurement of Tumor Necrosis Factor alpha (TNFa) in human serum, plasma, cell culture supernatants, and lysate samples.

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Antigen Information

Accession Number:
  • P01375
Gene ID:
  • 7124
Gene Symbols:
  • TNF
  • TNFA
  • TNFSF2
Protein Name & Synonyms:
Tumor necrosis factor (Cachectin) (TNF-alpha) (Tumor necrosis factor ligand superfamily member 2) (TNF-a) [Cleaved into: Tumor necrosis factor, membrane form (N-terminal fragment) (NTF) Intracellular domain 1 (ICD1) Intracellular domain 2 (ICD2) C-domain 1 C-domain 2 Tumor necrosis factor, soluble form]
Target Species:
  • Human

Assay Format

Detection Range:
30 pg/ml - 6000 pg/ml
Recommended Dilution (serum & plasma):
2 fold
30 pg/ml

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This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human ANG, CD23, Eotaxin, GCSF, GM-CSF, GRO-alpha, GRO-beta, GRO-gamma, I-309, IFN-gamma, IL-1 alpha, IL-1 beta, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p40), IL-12 (p70), IL-15, IL-16, IP-10, MCP-1, MCP-2, MCP-3, MCP-4, MCSF, MIG, MIP-1 alpha, MIP-1 beta, NAP-2, PDGF, PF-4
Number of Targets Detected:
Species Detected:
  • Human
Compatible Sample Types:
  • Cell Culture Supernatants
  • Cell Lysates
  • Plasma
  • Serum
  • Tissue Lysates
Design Principle:
  • Sandwich-based
Method of Detection:
  • Quantitative
Solid Support:
96-well Microplate

Product Specifications

1, 2, or 5 x 96-Well Strip Microplate Kit

Product Features

Additional Notes
  • Strip plates and additional reagents allow for use in multiple experiments
  • Quantitative protein detection
  • Establishes normal range
  • The best products for confirmation of antibody array data

Standard Curves

Standard curve graphs showing the results from the human tnf alpha elisa kit

Dilution series diagram with 8 vials

Spiking & Recovery Results

Sample Type Average % Recovery Range (%)
Serum 94.53 82-103
Plasma 91.92 80-102
Cell culture media 96.43 84-104

Linearity Results

    Serum Plasma Cell Culture Media
1:2 Average % of expected
Range (%)
1:4 Average % of expected
Range (%)

Application Notes

Kit Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Stop Solution
  • Assay Diluent(s)
  • Lyophilized Standard
  • Biotinylated Detection Antibody
  • Streptavidin-Conjugated HRP
  • TMB One-Step Substrate
Other Materials Required
  • Distilled or deionized water
  • Precision pipettes to deliver 2 µl to 1 µl volumes
  • Adjustable 1-25 µl pipettes for reagent preparation
  • 100 µl and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • Log-log graph paper or computer and software for ELISA data analysis
Protocol Outline
  1. Prepare all reagents, samples and standards as instructed in the manual.
  2. Add 100 µl of standard or sample to each well.
  3. Incubate 2.5 h at RT or O/N at 4°C.
  4. Add 100 µl of prepared biotin antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 µl of prepared Streptavidin solution to each well.
  7. Incubate 45 min at RT.
  8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 µl of Stop Solution to each well.
  11. Read at 450 nm immediately.

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The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
  1. Wu SJ, Lu TM, Lai MN, Ng LT. Immunomodulatory Activities of Medicinal Mushroom Grifola frondosa Extract and Its Bioactive Constituent. Am J Chin Med. 2013;41:131. doi: 10.1142/S0192415X13500109
    Species: Human
    Sample type: Conditioned Media
  2. Cervia, Davide, et al. "The protein pheromone Er-1 of the ciliate Euplotes raikovi stimulates human T-cell activity: involvement of interleukin-2 system."?Experimental cell research?319.3 (2013): 56-67.
    Species: Human
    Sample type: Conditioned Media
  3. Tian, Wenfang, et al. "Brahma?related gene 1 bridges epigenetic regulation of proinflammatory cytokine production to steatohepatitis in mice."?Hepatology58.2 (2013): 576-588.
    Species: Human
    Sample type: Conditioned Media
  4. Massicot, France, et al. "P2X7 cell death receptor activation and mitochondrial impairment in oxaliplatin-induced apoptosis and neuronal injury: cellular mechanisms and in vivo approach."?PLoS One?8.6 (2013): e66830.
    Species: Human
    Sample type: Conditioned Media
  5. Hinojosa, Ara E., et al. "Dual effects of noradrenaline on astroglial production of chemokines and pro-inflammatory mediators."?Journal of neuroinflammation?10.1 (2013): 852.
    Species: Human
    Sample type: Conditioned Media
  6. Chatterjee, Priyajit, et al. "Adipocyte fetuin-A contributes to macrophage migration into adipose tissue and polarization of macrophages."?Journal of Biological Chemistry?288.39 (2013): 28324-28330.
    Species: Human
    Sample type: Conditioned Media
  7. Zahran, S. M., et al. "Study of host immunity in patients with Helicobacter pylori-related idiopathic thrombocytopenia."?Eastern Mediterranean Health Journal?19.7 (2013).
    Species: Human
    Sample type: Serum
  8. Fathy, Mona Mohamad, et al. "TNF-?-308 promoter G/A and PTPN22 (1858 C/T) genes polymorphisms in Egyptian patients with systemic lupus erythematosus."?Comparative Clinical Pathology?22.5 (2013): 947-954.
    Species: Human
    Sample type: Serum
  9. Del Coco, Laura, et al. "A proton nuclear magnetic resonance-based metabolomic approach in IgA nephropathy urinary profiles."?Metabolomics?9.3 (2013): 740-751.
    Species: Human
    Sample type: Serum
  10. Badr, Rawia I., et al. "Serum sFas, TNF-? and Bcl 2 Expression Following Traumatic Brain Injury."?The Egyptian Journal of Medical Microbiology?38.1233 (2013): 1-12.
    Species: Human
    Sample type: Serum
Worked great with THP-1 cells stimulated with LPS. Sensitive and easy to use.

Duke University
Great kit. Easy to follow protocol. Getting great results.

Florida A&M University
It's so easy to use. I'm happy with it and recommend it to everyone.

Columbia University
Simple protocol and reproducible results. Would recommend.

University of Houston
I used it for human corneal epithelial cell culture supernatant under induced conditions. It is very sensitive and easy to use.

I also want to comment on the fast delivery of the product.

University of Illinois at Chicago
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This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.

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