Mouse Tumor Necrosis Factor-alpha, TNF-α HiLA Kit

Catalog No.: BHK1016

Size: 100 T

Detection Range: 0 pg/ml - 30,000 pg/ml

Linear Range: 2.5 pg/ml - 3,000 pg/ml

Storage and Expiration: Store at 2-8℃ in dark for 12 months.

Application: For quantitative detection of mouse TNF-α in buffer solution, serum, cell culture supernatant.

Introduction

Tumor necrosis factor (TNF), formerly known as TNF-α, is a chemical messenger produced by the immune system that induces inflammation. TNF is produced primarily by activated macrophages, and induces inflammation by binding to its receptors on other cells. It is a member of the tumor necrosis factor superfamily, a family of transmembrane proteins that are cytokines, chemical messengers of the immune system. Excessive production of TNF plays a critical role in several inflammatory diseases, and TNF-blocking drugs are often employed to treat these diseases.

Principle of the Assay  

Incubate the acceptor beads coated with anti- TNF-α antibody 1, biotin conjugated anti- TNF-α antibody 2, and the sample to form a sandwich immune complex (antibody 1-sample-antibody 2). The immune complex is then incubated with donor beads coated with streptavidin to form a chemiluminescent complex, ensuring that the distance between the two beads is less than 200 nm. When the distance between the two beads is less than 200 nm, the donor beads is excited by light (680 nm), producing singlet oxygen which diffuses to the acceptor beads, inducing energy transfer. The acceptor beads absorb this energy and emit scattering light at 615 nm. The light signal is collected by a photosensitive detector, and the concentration of TNF-α in the sample is calculated through mathematical fitting. In the absence of human TNF-α in the sample, no immune complex is formed, or the distance between the two beads exceeds 200 nm, exceeding the transfer distance for singlet oxygen, and the acceptor beads do not emit any light.

Kit components

Material Required But Not Provided

1. Deionized water
2. Luminescence plate or strip
3. Homogeneous chemiluminescence analyzer, or multimode microplate reader with Alpha module

Precautions

1. It is recommended to aliquot the reconstituted standard and store at -20℃, the series of gradient standards should be used within 2 h. Avoid repeated freeze-thaw cycles.
2. It is recommended that the dilution matrix for the standard be consistent with the dilution matrix of the sample to be tested.
3. Do not use the expired components and the components from different batches.
4. For your safety and health, please wear the lab coat and disposable gloves to operate.

Reference

1. van Loo G, Bertrand MJ (May 2023). "Death by TNF: a road to inflammation". Nature Reviews Immunology. 23 (5): 289–303.
2. Croft M, Siegel RM (March 2017). "Beyond TNF: TNF superfamily cytokines as targets for the treatment of rheumatic diseases". Nature Reviews Rheumatology. 13 (4): 217–233.
3. Jang DI, Lee AH, Shin HY, Song HR, Park JH, Kang TB, et al. (March 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.