Human TNF-alpha ELISA

The Human TNF alpha ELISA kit measures Tumor Necrosis Factor alpha (TNFa) in human serum, plasma, cell culture supernatants, and lysate samples. It contains a pre-coated 96-well strip plate with additional reagents suitable for multiple experiments.

$402.00  
ELH-TNFa-1
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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
Sensitivity:
30 pg/ml

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Specificity:
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:
1
Species Detected:
  • Human
Compatible Sample Types:
  • Cell Culture Supernatants
  • Cell Lysates
  • Plasma
  • Serum
  • Tissue Lysates
Design Principle:
  • Sandwich-based
Method of Detection:
Colorimetric
Quantitative/Semi-Quantitative:
  • Quantitative
Solid Support:
96-well Microplate

Product Specifications

Size:
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

Spiking and recovery results from the human tnf alpha elisa kit

Linearity Results

Linearity results for the human tnf alpha elisa kit

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.

Storage/Stability

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. Al Bahrawy, Mohammed M., et al. "The mutual effect of hyperlipidemia and proinflammatory cytokine related to periodontal infection."?Egyptian Journal of Oral and Maxillofacial Surgery?2.2 (2011): 87-95.
    Species: Human
    Sample type: Serum
  2. Kormann, Michael SD, et al. "Expression of therapeutic proteins after delivery of chemically modified mRNA in mice."?Nature biotechnology?29.2 (2011): 154-157.
    Species: Human
    Sample type: Conditioned Media
  3. Elsayeh, H., et al. "Study of anticardiolipin antibody in hepatitis C virus-positive patients."?Journal of Venomous Animals and Toxins including Tropical Diseases?17.4 (2011): 467-472.
    Species: Human
    Sample type: Serum
  4. Perez-Garcia JL., et al. Identification of TNF-a and MMP-9 as potential baseline predictive serum markers of sunitinib activity in patients with renal cell carcinoma using a human cytokine array. British Journal of Cancer (2009) 101, 1876 - 1883
    Species: Human
    Sample type: Serum
  5. Wu S-J, No L-T. Tetrandrine Inhibits Proinflammatory Cytokines, iNOS and COX-2 Expression in Human Monocytic Cells. Biol Pharm Bull. 2007;30:59-62.
    Species: Human
    Sample type: Conditioned Media
  6. Filková M, Hulejová H, Kuncová K, et al. Resistin in idiopathic inflammatory myopathies. Arthritis Research & Therapy. 2012;14(3):R111. doi:10.1186/ar3836.
    Species: Human
    Sample type: Serum
  7. Cerezo, Lucie Andrés, et al. "The metastasis promoting protein S100A4 is increased in idiopathic inflammatory myopathies."?Rheumatology?50.10 (2011): 1766-1772.
    Species: Human
    Sample type: Conditioned Media
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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Usage

This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.

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