This IFN-γ enzyme-linked immunosorbent assay (ELISA) applies a technique called a quantitative sandwich immunoassay. The microtiter plate provided in this kit has been pre-coated with a monoclonal specific for IFN-γ. Standards or samples are then added to the appropriate microtiter plate wells and incubated. IFN-γ, if present, will bind and become immobilized by the antibody pre-coated on the wells. The microtiter plate wells are thoroughly washed to remove unbound IFN-γ and other components of sample. In order to quantitatively determine the amount of IFN-γ present in the sample, a standardized preparation of horseradish peroxidase (HRP)-conjugated monoclonal antibody specific for IFN-γ is added to each well to "sandwich" the IFN-γ immobilized during the first incubation. The microtiter plate then undergoes a second incubation. The wells are thoroughly washed to remove all unbound HRP-conjugated antibodies and a TMB (3,3'5, 5' tetramethyl-benzidine) substrate solution is added to each well. The enzyme (HRP) and substrate are allowed to react over a short incubation period. Only those wells that contain IFN-γ and enzyme-conjugated antibody will exhibit a change in colour. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the colour change is measured spectrophotometrically at a wavelength of 450nm ± 2nm. In order to measure the concentration of IFN-γ in the samples this kit contains two calibration diluents. (Calibrator Diluent I for serum/plasma testing and Calibrator Diluent II for cell culture supernatant/ urine testing). According to the testing system, the provided standard is diluted (2-fold) with the appropriate Calibrator Diluent and assayed at the same time as the samples. This allows the operator to produce a standard curve of Optical Density (O.D.) versus IFN-γ concentration (pg/mL). The concentration of IFN-γ in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Platform
Microplate
Detection Type
Colorimetric
Sample Type
Serum, plasma, cell culture supernatant, and other biological fluids.
Sensitivity
< 7.5 pg/ml
Product Range
7.5-1600 pg/ml
Assay Time
2h 30m
Specificity
This IFN-γ ELISA Kit recognizes both natural and recombinant human IFN-γ.
Reactivity
Human
Cross Reactivity
This assay has shown no detectable cross-reactivity with human; SAA, EGF, IL-6, IL-8, IL-16, M-CSF, GM-CSF, TGF-β1, RANTES, FGF, MIP-1α, CRP, EPO.
Storage
Store at 2-8°C.
Product Note
Information online is representative. Always refer to the Product Manual inside the kit.
Disclaimer
This product is for research use only. It is not intended for diagnostic or therapeutic use.
