[Study on the properties of solid substrate room temperature phosphorescence of the antibody labeled with fluorescein isothiocynate].
The properties of room temperature phosphorescence (RTP) of the labeled goat anti-human antibody(with fluorescein isothiocynate GAHAb-FITC) and labeled rabit anti-goat antibody(RAGAb-FITC) with FITC in different dilution ratios on many kinds of solid substrates were studied in detail. It is found that on the polyamide membrane, only using Pb(Ac)2 as a heavy atom perturber, both the labeled antibodies and their antigen-antibody conjugated compound with the human immunoglobulin G remained excellent RTP properties of isothiocyanate fluorescein, lambda ex(max)/lambda em(max) = 525/650 nm, the relation of RTP intensity to the concentration was linear in a certain range. The conditions for immunological reaction and RTP emission were optimized. The result means that it is possible to develop a new immunoassay named as solid substrate room temperature phosphorescence immunoassay (SS-RTP-IA), which combines both the specificity of immunological reaction and high sensitivity of luminescence analysis.
Lui
Jia-ming
JM
Department of Chemistry, Tsinghua University, Beijing 100084, China.
Fu
Yan
Y
Yu
Bing-bin
BB
Li
Long-di
LD
D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D000888:Antibodies, Anti-Idiotypic / Q000737:chemistry* / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000737:chemistry / Q000276:immunology; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006041:Goats; D006801:Humans; D007118:Immunoassay / Q000379:methods; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology*; D049449:Luminescence; D011817:Rabbits; D013696:Temperature
D000906:Antibodies; D000888:Antibodies, Anti-Idiotypic; D000936:Antigen-Antibody Complex; D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D016650:Fluorescein-5-isothiocyanate