Human Tumor Necrosis Factor-beta, TNF-β HiLA Kit

Catalog No.: BHK1017

Size: 100 T

Detection Range: 0 pg/ml - 100,000 pg/ml

Linear Range: 0.35 pg/ml - 3,000 pg/ml

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

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

Introduction

Lymphotoxin-alpha (LT-α) formerly known as tumor necrosis factor-beta (TNF-β) is a protein that in humans is encoded by the LTA gene. Belonging to the hematopoietic cell line, LT-α exhibits anti-proliferative activity and causes the cellular destruction of tumor cell lines. As a cytotoxic protein, LT-α performs a variety of important roles in immune regulation depending on the form that it is secreted as. Unlike other members of the TNF superfamily, LT-α is only found as a soluble homotrimer, when found at the cell surface it is found only as a heterotrimer with LTβ.

Principle of the Assay  

Incubate the F conjugated anti- TNF-β antibody 1, biotin conjugated anti- TNF-β antibody 2, acceptor beads coated with F-conjugate, 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-β in the sample is calculated through mathematical fitting. In the absence of human TNF-β 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. Calmon-Hamaty F, Combe B, Hahne M, Morel J (2011). "Lymphotoxin α revisited: general features and implications in rheumatoid arthritis". Arthritis Research & Therapy. 13 (4): 232.
2. Nedwin GE, Naylor SL, Sakaguchi AY, Smith D, Jarrett-Nedwin J, Pennica D, et al. (September 1985). "Human lymphotoxin and tumor necrosis factor genes: structure, homology and chromosomal localization". Nucleic Acids Research. 13 (17): 6361–6373.
3. Aggarwal BB, Eessalu TE, Hass PE (February 1986). "Characterization of receptors for human tumour necrosis factor and their regulation by gamma-interferon". Nature. 318 (6047): 665–667.