Peroxidase-antiperoxidase method

S.M. Hsu, L. Raine, and H. Fanger, A comparative study of the peroxidase-antiperoxidase method and an avidin-biotin complex method for studying polypeptide hormones with radioimmunoassay antibodies. Am. J. Clin. Pathol 75, 734-738 (1981). 

Fig. 1. Schematic drawing of various immune methods for localization of antigens. (A) Direct method. (B) Indirect method. (C) Immunogold with silver enhancement. (D) Peroxidase-antiperoxidase method. (E) Avidin-biotin system.

Key words Direct assay, Indirect assay, Peroxidase-antiperoxidase method, Immune polymer, DAB chromagen, PBS buffer, TBS buffer... 

Fig. 1, from 7 days before the partial pancreatectomy, nicotinamide, at a dose of 0.5 g kg" body weight in 2 ml saline, and 3-aminobenzamide HCl, at a dose of 0.05 g kg" body weight in 2 ml saline, were injected intraperitoneally into each group of five rats every day. Nicotinamide and 3-aminobenzamide are inhibitors of pancreatic B-cell poly(ADP-ribose) synthetase the inhibitory ability of the latter is about tenfold as potent as that of the former [9]. The injection was continued until the 90th postoperative day. Five control rats received saline daily before and after the partial pancreatectomy. The residual pancreatic tissue, referred to as the remaining pancreas, was removed 3 months after the partial pancreatectomy and fixed in Bouin s solution. Hydrated 5 juni sections of paraffin-embedded pancreatic tissues were stained for insulin, glucagon, and somatostatin by the peroxidase antiperoxidase method [14]. 

The Peroxidase-Antiperoxidase (PAP) Method and Other All-Immunologic Detection Methods ... 

Elias, J., Margiotta, M., and Gabore, D. (1989) Sensitivity and detection efficiency of the peroxidase antiperoxidase (PAP), avidin-biotin complex (ABC), and the peroxidase-labeled avidin-biotin (LAB) methods. Am. J. Clin. Pathol. 92,62. 

The avidin-biotin complex (ABC) method and the streptavidin-biotin (SAB) method are more sensitive than the peroxidase-antiperoxidase (PAP) method for histochemical techniques. The strong noncovalent attraction between biotin and avidin or streptavidin is exploited in many histochemical, immunohistochemical, and in situ hybridization... 

An immunohistochemical examination of PSA using polyclonal antibodies by the peroxidase antiperoxidase (PAP) method and by the technique of biotin-streptavidin-alkaline phosphatase has been successfully carried out (Zaviacic et al., 1994). Immunoelectron microscopy in conjunction with the protein A-gold complex can also be used for localizing PSA in human prostate (Sinha et al., 1987). The procedure for immunofluorescence localization of PSA is given below. 

A variety of techniques (Fig. 1), which use a variable combination of antibody steps, enzyme labels, and chromogens, are employed in immunohistochemistry. The most common immunohistochemical methods are the PAP (peroxidase-antiperoxidase), ABC (avidin-biotin) peroxidase or alkaline phosphatase, and the APAAP (alkaline phosphatase-anti-alkaline phosophatase) techniques however,... 

In the second version, the separate application of anti-PO and PO was combined into a single step. For this purpose soluble peroxidase-antiperoxidase PO-(rabbit)-anti-PO has been prepared. Details can be found elsewhere This method is one example of labeling in which covalent binding is not necessary. 

Fig. 12. Diagram showing sequence for detecting viral antibody by the peroxidase-antiperoxidase (PAP) method of Stemberger et al. This method uses a bridging antibody (anti-primate antibody) between the human and simian antibody. Artificial (chemical) linkage of enzyme with antibody is replaced by an antigen-antibody reaction.

Lindskog S, Friskopp J Immunoglobulins in human dental calculus demonstrated with the peroxidase-antiperoxidase (PAP) method. Scand J Dent Res 1983 91 360-364. 

Kiviluoto M, Rasanen 0, Rinne A, et al. 1981. Intracellular immunoglobulins in plasma cells of nasal biopsies taken from vanadium-exposed workers A retrospective case control study by the peroxidase-antiperoxidase (PAP) method. Anat Anz 149 446-450. 

Sternberger LA, Sternberger NH (1986) The unlabeled antibody method comparison of peroxidase-antiperoxidase with avidin-biotin complex by a new method of quantification. J Histochem Cytochem 34 599-605... 

FIGURE 1.6 Enzyme bridge method. A second antibody is used to link (bridge) the primary antibody to an antiperoxidase antibody, which, in turn binds to free peroxidase. Boxed asterisk represents antigen determinant on primary and secondary antibodies. Px, peroxidase label. From Taylor CR, Cote RJ. Immunomicroscopy A Diagnostic Tool for the Surgical Pathologist. 2nd ed. Philadelphia WB Saunders 1994 12. 

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