A new procedure is described for extracting, purifying, and crystallizing alkaline phosphatase orthophosphoric monoester phosphohydrolase (EC 3.1.3.1) from human intestinal mucosa. Active enzyme, extracted with trichiorotrifluoroethane, is subsequently purified by chromatography on Sephadex G-200 and DEAE Sephadex. The acetone and ammonium sulfate precipitation steps that are part of the conven...

The action of non-detergent sulphobetaines (NDSBs) as new mild agents for protein purification is described. The solubilization effects of non-detergent sulphobetaines are shown in different examples; all obtained under non-denaturing conditions: (1) microsomal proteins extraction; (2) recovery after dialysis of nuclear proteins; (3) reduction of precipitation in isoelectric focusing experiment...

Two enzyme forms of alkaline phosphatase have been partially purified from the medium spent for the culture of HUH-6 clone 5 cells, which were originally derived from hepatoblastoma tissue. The purification methods used are ammonium sulfate precipitation, ethanol precipitation, diethylaminoethyl cellulose chromatography, Affi-Gel Blue chromatography, and Sephadex G-200 gel filtration. These alk...

We developed a new simple high-throughput plasmid DNA extraction procedure, based on a modified alkaline lysis method, using only one 96-well microtiter glassfilter plate. In this method, cell harvesting, lysis by alkaline and plasmid purification are performed on only one microtiter glassfilter plate. After washing out RNAs or other contaminants, plasmid DNA is eluted by low-ion strength solut...

BACKGROUND Too many different protein and enzyme purification techniques have been reported, especially, chromatographic techniques. Apart from low recovery, these multi-step methods are complicated, time consuming, high operating cost. So, alternative beneficially methods are still required. Since, the outstanding advantages of aqueous two phase system (ATPS) such as simple, low cost, high rec...

INTRODUCTION..........................................155 METHODS FOR DETECTING MULTIPLE MOLECULAR FORMS OF AN ENZYME.............155 HEXOKINASE...........................................157 MULTIPLE FORMS OF ENZYMES APPEARING As ARTIFACTS OF PURIFICATION............158 LACTIC DEHYDROGENASE....................................158 ALKALINE PHOSPHATASE....................................161 ESTERA...

This report relates to the preceding paper by Taniuchi and Anfinsen. Micrococcal nuclease from the strain “Foggi” has been purified from a commercially available source (Worthington). The enzyme was crystallized from an ammonium sulfate solution. The over-all purification was approximately SOO-fold with an over-all yield of about 16%. The enzyme is virtually devoid of acid and alkaline phosphat...