Sensitivity and Detection Efficiency of the Peroxidase Antiperoxidase (PAP), Avidin–Biotin Peroxidase Complex (ABC), and Peroxidase-Labeled Avidin–Biotin (LAB) Methods

Abstract

The authors have examined the sensitivity and detection efficiency of the three peroxidase methods that currently have the widest application in diagnostic immunohistochemistry: the peroxidase-antiperoxidase (PAP), the avidin-biotin complex (ABC), and the labeled avidin-biotin (LAB) methods. Sensitivity was evaluated by determining the highest useful dilution of polyclonal antiglucagon antibodies applied to formalin-fixed, paraffin-embedded human pancreas. Detection efficiency was evaluated by tabulation of the total number of positive (three or more positive cells) islets. On direct comparison, the LAB method exceeded the PAP and ABC methods in both sensitivity and detection efficiency, which were essentially equal. Titration of linking antiserum of the PAP method boosted its sensitivity and detection efficiency above that of ABC; the PAP had equal sensitivity to the LAB and exceeded it in detection efficiency. The authors conclude that comparisons of immunohistologic methods are meaningful only if both sensitivity and efficiency are considered along with the unique requirements of any single method.

Down-Regulation of CD9 Expression during Prostate Carcinoma Progression Is Associated with CD9 mRNA Modifications

“…Endogenous peroxidase was blocked by 1.5% of hydrogen peroxide in 50% methanol. An immunohistochemistry kit, HistostainTM-SP kit (Zymed, South San Francisco, CA), was used to detect CD9 signals (32). Tissues were first incubated with nonimmune goat serum for 25 min to minimize nonspecific binding and then incubated with monoclonal antihuman CD9 mouse antibody (NCL-CD9; dilution of 1:500) for 1 h. The CD9 antibody recognizes an epitope located in the extracellular region of the protein.…”

Detection of human parvovirus B19 in papillary thyroid carcinoma

“…Then the sections were incubated overnight at 41C with the mouse monoclonal antibody against the B19 proteins VP1/VP2 (1 : 20 dilution; Novocastra, Newcastle, UK) and the mouse monoclonal antibody against the nuclear transcription factor subset NF-kB p65 (1 : 400 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA), respectively. A streptavidin -biotin -peroxidase complex kit (SABC kit, Zymed, South San Francisco, CA, USA) was used to detect the protein conjugates (Elias et al, 1989). Finally, the sections were developed with a diaminobenzidine substrate (Sigma, St Louis, MO, USA) and counterstained with haematoxylin.…”

Rapid diagnosis of infectious laryngotracheitis using a monoclonal antibody‐based immunoperoxidase procedure

“…A variety of IP procedures using unlabeled or peroxidase-labeled antibodies have been described (Hsu et al, 1981;Shi et al, 1988;Elias et al, 1989). The streptavidinperoxidase conjugate (SP) IP procedure utilized in this study is a modification of the avidin-biotin-peroxidase complex (ABC) procedure described by Hsu et al (1981).…”

“…The SP IP procedure utilizes unlabeled primary antibody, a biotinylated secondary antibody, and a streptavidin-peroxidase conjugate. Recently, this IP procedure was shown to provide greater sensitivity than other commonly used IP procedures including ABC and peroxidase-antiperoxidase techniques (Shi et al, 1988;Elias et al, 1989).…”