Localization of Prostate-Specific Antigen

Prostate-specific antigen is currently the most frequently used marker for identifying normal and pathologically altered prostatic tissue in the male and female. At present, there 

A healthy female with a normal prostate is characterized by a broad range of serum PSA values from practically unappreciable amounts to the highest reported ones of 0.9 ng/ml. This value is very close to the normal reference range in the male (Zaviadic and Ablin, 2000). 

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. 

Paraffin-embedded prostatic tissue sections (5 xm thick) are deparaffinized and rehydrated (Scorilas et al., 2000). A 5% universal tissue conditioner (Biomeda) is applied for 10 min at room temperature to block any nonspecific binding. The sections are incubated with biotinylated monoclonal mouse antihuman PSA antibody ( 2mg/l) (Diagnostic Systems Laboratories [coded 8301]) for 1 hr at 37°C. The sections are stained with (SA)Z-(biotin)x-PVA-(BCPDA)y-Eu3+ for 25 min at 37°C. After each step, the sections are rinsed briefly with 0.5 ml/1 Tween 20 solution. The slides are dried with a stream of cold air, and the resultant fluorescence is observed with a time-resolved fluorescence microscope. 

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Sinha, A. A., Wilson, M. J., and Gleason, D. F. 1987. Immunoelectron microscopic localization of prostatic-specific antigen in human prostate by the protein A-gold complex. Cancer 60 1288-1293. 

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