To establish a highly sensitive time-resolved immunofluorometric assay of human interferon-gamma. A two-site “Sandwich”-type time-resolved immunoflourometric assay for human interferon-gamma is described. It is based on usage of specific polyclonal and monoclonal antibodies. The polyclonal antibody to bind the interferon-gamma in samples are immoblailized on the surface of microtiter plate strip wells.Following capture of IFN-γ present in sample. Then monoclonal antibody, which is biotinylated,is added to wells.The second biotin labeled antibody will allow the subsequent specific binding of Eu3+ labeled streptavidin. After the immunoreactions completed, the bound fraction of Eu3+-label is quantified by dissociating it in a fluorescence-enhancement solution and measuring its fluorescence with Wallac 1234 fluorometer. The sensitivity of the assay is 0.02 μg/L. The standard curve is linear from 0.02 μg/L to 400 μg/L. The time-resolved immunofluorometric IFN-γ assay is quick, sensitive and suitable for testing large numbers of samples, and may be useful in both industrial producing and clinical studies.
Erdeni, GUO Mei-Xiang, HAN Ling, QI Qi-Ge, LIU Shi-Shan, ZHANG Li-Min, CHEN Qi, ZHANG Qing-Rong. Two-site Time-resolved Immunofluorometric Assay of Human Interferon-gamma[J]. Progress in Biochemistry and Biophysics,1999,26(4):388-391
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