Reducing non-specific background in immunofluorescence technique

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...

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435 5, 436511, 436519, 436548, 436800, 436825, 25230116, G01N 5300, G01N 33571, G01N 33554, G01N 3353

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045827915

ABSTRACT:
Novel compositions and methods are provided for detecting the presence of a material of interest in a specimen on a solid surface. A composition useful for detecting the presence of a material of interest in a specimen comprises (1) a detector conjugate comprising a fluorescing moiety bonded to a compound capable of specific binding with the material of interest, or a derivative thereof, and (2) a non-detector conjugate comprising a poly(amino acid) and a compound having substantial structural and charge similarity to the fluorescing compound and no observable fluorescence, or low level fluorescence of a different wavelength than that of the fluoresing compound, in the region of fluorescence of the fluorescing compound. In the method of the invention, a specimen, on a solid surface, is combined with the detector conjugate and the non-detector conjugate, and the combination is incubated. Unbound conjugates are removed from the solid surface by washing and then the surface is irradiated with light having a wave length absorbed by the fluorescing compound. The fluorescent light emitted by the combination is observed and is a function of the amount of the material of interest present in the specimen.

REFERENCES:
patent: 4208479 (1980-06-01), Zuk et al.
patent: 4261968 (1981-04-01), Ullman et al.
patent: 4351760 (1982-09-01), Khanna et al.
Techniques in Immunocytochemistry, vol. 1, (1982) K. B. Pryzwansky, pp. 77-89, Academic Press, Pubs.

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