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KMID : 0357919740080020131
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Korean Journal of Pathology
1974 Volume.8 No. 2 p.131 ~ p.137
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Simple Microzone Cellulose Acetate Electrophoresis of Alkaline Phosphatase Iscenzymes in Human Tissue and Serum
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김재식/Jae Sik Kim
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Abstract
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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 thev
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 characterizatioin 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).
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