Chromatographic separation of alkaline phosphatase from dental enamel.
Alkaline phosphatase (AP) was prepared from partly mineralized bovine enamel by extraction in phosphate buffer, centrifugation and various chromatographic techniques. Chromatofocusing showed that the enamel enzyme possessed five isoelectric points at the acid pH level ranging from pH 5.7 to pH 4.4. Three enzyme peaks were eluted using low pressure chromatography with a Bio-gel column. With a HPLC gel filtration column the separation of the enamel extract resulted in only one peak with AP activity. The fractions of this peak were used to produce an antibody against bovine AP.
Moe
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D
Institutes of Biochemistry, Royal Dental College, Copenhagen, Denmark.
Kirkeby
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S
Salling
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E
D000469:Alkaline Phosphatase / Q000032:analysis*; D000818:Animals; D002417:Cattle; D002845:Chromatography; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003743:Dental Enamel / Q000032:analysis / Q000201:enzymology*; D003746:Dental Enamel Proteins / Q000032:analysis; D004586:Electrophoresis; D007525:Isoelectric Focusing; D013053:Spectrophotometry
D003746:Dental Enamel Proteins; D000469:Alkaline Phosphatase