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Human TNF-alpha ELISA

RayBio® Human TNF alpha ELISA Kit for cell culture supernatants, plasma, serum, 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

Spiking & Recovery Results

Linearity Results

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.


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. Wang P., Ma X., He Y., et al. Effect of p38 mitogen-activate protein kinase on MUC5AC protein expression of bile duct epithelial cells in hepatolithiasis patients.
    Species: Human
    Sample type: Conditioned Media
    Species: Human
    Sample type: Serum
  3. Bishayi B., et al. Expression of CXCR1 (Interleukin-8 Receptor) in Murine Macrophages After Staphylococcus aureus Infection and its Possible Implication on Intracellular Survival Correlating with Cytokines and Bacterial Anti-Oxidant Enzymes. Inflammation August 2014, Epub ahead of print. DOI: 10.1007/s10753-014-9991-1
    Species: Human
    Sample type: Conditioned Media
  4. Chhabra G.S., Kulkarni K.S. Evaluation of anti-inflammatory activity of Vitex agnus castus leaves. Quantitative analysis of flavonoids as possible active constituents. ~?183 ~Journal of Pharmacognosy and Phytochemistry 2014; 3 (1): 183-189
    Species: Rat
    Sample type: Serum
  5. Duan J, Yu Y, Yu Y, et al. Silica nanoparticles induce autophagy and endothelial dysfunction via the PI3K/Akt/mTOR signaling pathway. International Journal of Nanomedicine 2014;9:5131-5141. doi:10.2147/IJN.S71074.
    Species: Human
    Sample type: Conditioned Media
  6. Gao, S., Dine, J., et al. Anti-inflammatory and Anti-apoptotic Effect of Combined Treatment with Methylprednisolone and Amniotic Membrane Mesenchymal Stem Cells After Spinal Cord Injury in Rats. Neurochemical Research 2014. Epub Ahead of Print DOI:10.1007/s11064-014-1344-9
    Species: Human
    Sample type: Tissue Lysate
  7. Gheita T., Azkalany G., Gaber E., Mohey A. Clinical significance of serum TNF? and -308 G/A promoter polymorphism in rheumatoid arthritis. The Egyptian Rheumatologist (2014), Epub Ahead of Print. http://dx.doi.org/10.1016/j.ejr.2014.07.001
    Species: Human
    Sample type: Serum
  8. Rattray B., Steel J., Semple S., Ramirez R., Abhayaratna WP. Short-term eccentric exercise in newly diagnosed type II diabetics: an exploratory study. Sport Sciences for Health, June 2014 Epub ahead of print
    Species: Human
    Sample type: Plasma
  9. Shaker MJ., et al. Immunological Study of Women Infected with Trichomonas vaginalis Parasite in Baghdad city. International Journal of Science and Research. Volume 3 Issue 9, September 2014,
    Species: Human
    Sample type: Serum
  10. Moore Q., Smith C., Wolf S., et al. Examining the immune response of pneumolysin exposed human corneal epithelial cells in a keratitis model. BIOS 85(4):199-206. 2014. doi: http://dx.doi.org/10.1893/0005-3155-85.4.199
    Species: Human
    Sample type: Conditioned Media
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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