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Human TNF-alpha ELISA

RayBio® Human TNF alpha ELISA Kit for cell culture supernatants, plasma, serum, and lysate samples.

$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

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.

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. Gheita T., Azkalany G., Gaber W., Mahoy A. Clinical significance of serum TNFa and -308 G/A promoter polymorphism in rheumatoid arthritis. The Egyptian Rheumatologist, In Press Jyly 2104. http://dx.doi.org/10.1016/j.ejr.2014.07.001
    Species: Human
    Sample type: Serum
  2. Deangelo DJ, Mesa RA, Fiskus W, et al. Phase II trial of panobinostat, an oral pan-deacetylase inhibitor in patients with primary myelofibrosis, post-essential thrombocythaemia, and post-polycythaemia vera myelofibrosis. Br J Haematol. 2013 Aug;162(3):326-335.
    Species: Human
    Sample type: Plasma
  3. Ertugrul AS, Sahin H, Dikilitas A, et al. Comparison of CCL28, interleukin-8, interleukin-1? and tumor necrosis factor-alpha in subjects with gingivitis, chronic periodontitis and generalized aggressive periodontitis. J Periodont Res. 2013 Feb;48(1):44-51.
    Species: Human
    Sample type: Gingival Crevicular Fluid
  4. Fukumoto J, Fukumoto I, Parthasarathy PT, et al. NLRP3 Deletion Protects from Hyperoxia-Induced Acute Lung Injury. Am J Physiol Cell Physiol. 2013 Jul;305(2):C182-C189.
    Species: Mouse
    Sample type: BAL Fluid
  5. Hung P., Tsai H., Lin C., Hung K. Abdominal Obesity is Associated with Peripheral Artery Disease in Hemodialysis Patients. PLOS One Published: August 02, 2013 DOI: 10.1371/journal.pone.0069424
    Species: Human
    Sample type: Serum
  6. Leow K-Y, Goh WWB, Tan S-Z, et al. Variant screening of the serum amyloid A1 gene and functional study of the p.Gly90Asp variant for its role in atherosclerosis. Atherosclerosis. 2013 Mar;227(1):112-117.
    Species: Human
    Sample type: Conditioned Media
  7. Llamas-Covarrubias MA, Valle Y, Bucala R, et al. Macrophage migration inhibitory factor (MIF): Genetic evidence for participation in early onset and early stage rheumatoid arthritis. Cytokine. 2013;61(3):759-765.
    Species: Human
    Sample type: Serum
  8. Omoomi FD. Molecular Chlorambucil-Methionine Conjugate: Novel Anti-cancer Agent against Breast MCF-7 Cell Model. J Cancer Sci Therapy. 2013;05(02).
    Species: Human
    Sample type: Conditioned Media
  9. Rocher, Crystal, and Dinender K. Singla. "SMAD-PI3K-Akt-mTOR Pathway Mediates BMP-7 Polarization of Monocytes into M2 Macrophages." PLoS One. NCBI, 20 Dec. 2013. Web.
    Species: Human
    Sample type: Conditioned Media
  10. Verma P, Dalal K. Serum cartilage oligomeric matrix protein (COMP) in knee osteoarthritis: A novel diagnostic and prognostic biomarker. J Orthop Res. 2013 Jul;31(7):999-1006.
    Species: Human
    Sample type: Serum
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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