Biotin/avidin formulation

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

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435 72, 435 721, 435 79, 435960, 435962, 435967, 435975, G01N 33535

Patent

active

054879752

ABSTRACT:
The present invention provides an improved biotin-avidin formulation in which a biotinylated antibody conjugate or an avidin-enzyme conjugate is present in a suitable diluent for immunohistochemical staining. The diluent additionally comprises casein in an amount sufficient to prevent charge interactions of the conjugate with a tissue section and gamma globulin in an amount sufficient to prevent Fc receptor binding and any hydrophobic interaction of the conjugate with a tissue section. In a preferred embodiment, the immunoglobulin is from the same species as the biotinylated antibody conjugate. The formulation effectively reduces overall unwanted binding, irrespective of the source of the binding.

REFERENCES:
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patent: 5066581 (1991-11-01), Suciu-Foca et al.
patent: 5225325 (1993-07-01), Miller et al.
Duhamel, R. C. et al. "Prevention of nonspecific binding of avidin." Methods in Enzymology 184:201-207,1990.
Brandon, C., Improved Immunocytochemical Staining Through the Use of Fab Fragments of Primary Antibody, Fab-specific Second Antibody, and Fab-Horseradish Peroxidase, 33 The Journal of Histochemistry and Cytochemistry 715 (abstract) (1985).
Duhamel, R. and Johnson, D., Use of Nonfat Dry Milk to Block Nonspecific Nuclear and Membrane Staining by Avidin Conjugates, 33 The Journal of Histochemistry and Cytochemistry 711-714 (1985).
Hochhaus, G. and Sadee, W., A Biotin-Avidin-Based Enzyme Immunoassay for .beta..sub.h -Endorphin, 5 Pharmaceutical Research 232-235 (1988).

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