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  • Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures.

Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures.

The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society (1981-04-01)
S M Hsu, L Raine, H Fanger
ABSTRACT

The use of avidin-biotin interaction in immunoenzymatic techniques provides a simple and sensitive method to localize antigens in formalin-fixed tissues. Among the several staining procedures available, the ABC method, which involves an application of biotin-labeled secondary antibody followed by the addition of avidin-biotin-peroxidase complex, gives a superior result when compared to the unlabeled antibody method. The availability of biotin-binding sites in the complex is created by the incubation of a relative excess of avidin with biotin-labeled peroxidase. During formation of the complex, avidin acts as a bridge between biotin-labeled peroxidase molecules; and biotin-labeled peroxidase molecules, which contains several biotin moieties, serve as a link between the avidin molecules. Consequently, a "lattice" complex containing several peroxidase molecules is likely formed. Binding of this complex to the biotin moieties associated with secondary antibody results in a high staining intensity.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Biotin, ≥99.0% (T)
Sigma-Aldrich
Biotin, tested according to Ph. Eur.
Sigma-Aldrich
Anti-Avidin antibody produced in rabbit, whole antiserum
Sigma-Aldrich
Avidin from egg white, BioUltra, lyophilized powder, ≥10 units/mg protein (E1%/280), ≥98% (SDS-PAGE)
Sigma-Aldrich
Avidin–Alkaline Phosphatase, buffered aqueous solution
Sigma-Aldrich
Avidin–Peroxidase, lyophilized powder
Sigma-Aldrich
Avidin–FITC from egg white, buffered aqueous solution
Sigma-Aldrich
Avidin–FITC from egg white, lyophilized powder
Sigma-Aldrich
Avidin–Peroxidase, buffered aqueous solution