Abstract

The author attempted a newly applicable simple electrophoretic screening of alkaline phosphatase isoenzymes with human tissue extracts and serum on microzone cellulose membrane with multiple specimen. Isoenzymes were localized colorimetrically with they substrate alpha-naphthol AS-MX phosphate by coupling the reaction product with Fast blue RR salt and Fast red violet LB salt. Alkaline phosphatase consists of a cup of enzymes which hydrolyze phosphate esters in an alkaline medium as follows by the general reaction: R-O-P + H₂O AIPase P + ROH The increase of serum alkaline phosphatase activity is observed in hepatobiliary disease, various bone disorders characterized by increased osteoblastic activity, during growth, pregnancy and a certain kinds of intestinal tract disease etc. The alkaline phosphatase has been fractionated electrophoretically into isoenzymes by numerous methods including paper, starch ge1 cellulose acetate agar gel acrylamide disc gel cellogen micro starch gel and Beckman microzone agarose gel membrane because various electrophoretic media, substrates, buffers and conditions have been used to fractionate and localize isoenzymes of alkaline phosphatase (orthophosphoric monoester phosphohydrolase) from human serum and tissues, it is often difficult to compare results. Usually these methods were examined and found unsuitable for use in a routine clinical laboratory. The author’s objective was to devise a simple routine screening method for localization and characterization of alkaline phosphatase isoenzymes on microzone cellulose acetate membrane by the simplest procedure as well as microzon cellulose acetate electrophoresis of serum protein. Electrophoresis of eight samples on a single microzone cellulose membrane facilitates comparison of isoenzymes with known tissue extract controls(liver and bone).