Tumor necrosis factor alpha (TNF-α) is a cytokine that has pleiotropic effects on various cell types. It has been identified as a major regulator of inflammatory responses and is known to be involved in the pathogenesis of some inflammatory and autoimmune diseases. Structurally, TNF-α is a homotrimer protein consisting of 157 amino acids, mainly generated by activated macrophages, T-lymphocytes, and natural killer cells. It is functionally known to trigger a series of various inflammatory molecules, including other cytokines and chemokines.

Human TNF-alpha ELISA Kit Principle

This Human TNF-alpha ELISA Kit is based on sandwich ELISA, the TNF-alpha specific monoclonal antibody was pre-coated in the microwells, the TNF-alpha in sample or the TNF-alpha standard will be captured by the coated antibody, after wash step, the unbonded TNF-alpha will be removed, and then another TNF-alpha specific monoclonal antibody is added and incubated. After then, the plate is washed again, and the substrate is added. The color of the wells is positively proportional to the TNF-alpha in it. After adding the stop solution, OD values are measured with ELISA reader. Unknown samples can be calculated from the standard curve.

Kit Components

This Human TNF-alpha ELISA kit is a readily commercial product, it has the following components, which is sufficient for 42 testing in duplicated wells. Please always check the kit instruction manual for packaging details. For plasma and serum samples, the standard solutions are ready to use. For other samples, you may need to dilute the standard according to the kit instruction manual.

• Microtiter plate, 96wells,
• Standard, will be diluted to 7 vials,
• HRP conjugate, 1 vial,
• TMB substrate, 1 vial,
• Assay buffer, 1 vial,
• Wash buffer, 1 vial,
• Sealer film or plate cover, 1p pc
• Stop solution, 1 vial,

Specificity and sensitivity

The sensitivity is 0.21 – 0.26 pg/mL as MDD. The specificity is over 99%.

Result determination

The sample OD value is measured by an ELISA reader, then the standard curve is plotted. We recommend 4PL regression model or another equivalent model.

Storage and expiry

2-8 degree Celsius for at least 12 months.

Caution

• This kit is for research use only.
• The standard solution in the kit is human TNF-alpha.
• Please read the kit instruction manual carefully before testing.
• Each run will take 80min or more, please schedule your time before experiment.

Reference

1. Jang, D. I., Lee, A. H., Shin, H. Y., Song, H. R., Park, J. H., Kang, T. B., Lee, S. R., & Yang, S. H. (2021). The Role of Tumor Necrosis Factor Alpha (TNF-α) in Autoimmune Disease and Current TNF-α Inhibitors in Therapeutics. International journal of molecular sciences, 22(5), 2719. https://doi.org/10.3390/ijms22052719