New detection method for uropepsinogen (PGA) using isoelectric focusing and immunoblotting techniques

Abstract

Uropepsinogen (PGA) was isolated and purified from human urine using a column chromatography series. The purified PGA was injected into a rabbit and a PGA-specific antibody was obtained. PGA isozymogen in human urine could be detected reproducibly by immunoblotting using this antibody after isoelectric focusing electrophoresis (IEF) on polyacrylamide gels. This technique may prove to be useful in the genetic study of PGA polymorphism.

New detection method for ribonuclease 2 (RNase 2) using immunoblotting with specific antibody

“…After IEF-PAGE, the isozymes was transferred onto a Durapore strip (Millipore) essentially according to the method described in our previous papers [4][5][6][7][8]. In the case of SDS-PAGE, transfer was carried out by electroblotting using a KS-8453 apparatus (Marysol, Tokyo, Japan).…”

“…The combination of IEF-PAGE and immunoblotting with a specific antibody has been used as a valuable technique for investigating possible heterogeneity and new genetic polymorphisms of pepsinogen [4,5], deoxyribonuclease [6,7] and a urinary glycoprotein, GP 43 [8]. Accordingly, immunological detection of the secretory RNase in body fluids seems most suitable for genetic or forensic distinction between the two RNases coexisting in body fluids [1].…”

Salivary deoxyribounuclease I polymorphism separated by polyacrylamide gel‐isoelectric focusing and detected by the dried agarose film overlay method

“…These plasma samples were stored at -80°C until the DNase I phenotyping was performed. Urine samples were concentrated, dialyzed and finally lyophilized, as described previously [7,8]. A 0.1 Vo w/v solution of the lyophilized material, corresponding to tenfold concentrated urine, was used for DNase I phenotyping.…”