Human Interferon gamma, IFN-γ HiLA Kit

Catalog No.: BHK1018

Size: 100 T

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

Linear Range: 1.07 pg/ml - 10,000 pg/ml

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

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

Introduction

Interferon gamma (IFNG or IFN-γ) is a dimerized soluble cytokine that is the only member of the type II class of interferons. The existence of this interferon, which early in its history was known as immune interferon, was described by E. F. Wheelock as a product of human leukocytes stimulated with phytohemagglutinin, and by others as a product of antigen-stimulated lymphocytes. It was also shown to be produced in human lymphocytes. or tuberculin-sensitized mouse peritoneal lymphocytes challenged with Mantoux test (PPD); the resulting supernatants were shown to inhibit growth of vesicular stomatitis virus.

Principle of the Assay  

Incubate the acceptor beads coated with anti- IFN-γ antibody 1, biotin conjugated anti- IFN-γ 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 IFN-γ in the sample is calculated through mathematical fitting. In the absence of human IFN-γ 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. Gray PW, Goeddel DV (August 1982). "Structure of the human immune interferon gene". Nature. 298 (5877): 859–863
2. Wheelock EF (July 1965). "Interferon-Like Virus-Inhibitor Induced in Human Leukocytes by Phytohemagglutinin". Science. 149 (3681): 310–311.
3. Green JA, Cooperband SR, Kibrick S (June 1969). "Immune specific induction of interferon production in cultures of human blood lymphocytes". Science. 164 (3886): 1415–1417.
4. Milstone LM, Waksman BH (November 1970). "Release of virus inhibitor from tuberculin-sensitized peritoneal cells stimulated by antigen". Journal of Immunology. 105 (5): 1068–1071.