Kinetic determination of granulocyte alkaline phosphatase by the GEMSAEC. 1980

L Crook, and N M Sarji, and P I Liu, and R H Gadsden, and T R Williams

Granulocyte alkaline phosphatase (ALP) activity is measured kinetically using the GEMSAEC centrifugal analyzer and p-nitrophenylphosphate as substrate. Measurements of enzyme activity are made at pH 9.8 utilizing 1.0 M diethanolamine as a buffer containing 0.5 mM magnesium chloride. Granulocytes are separated from heparinized blood using dextran sedimentation, followed with ficoll-hypaque separation. The separated cells are suspended in saline, and the red cells are lysed. Manual counting of stained smears of the cells isolated showed that 95 percent of these cells were granulocytes. After lysing the red cells, the granulocyte suspension is rinsed in 12.5 percent heat inactivated serum in saline and separated by centrifugation. After 200 microliters of the diluent solution (12.5 percent heat inactivated serum in saline) is added to the cells, they are sonicated for seven sec at 0 degrees in an ice bath. The extracts are then assayed kinetically for ALP activity. Granulocyte ALP activity varies from day to day in normal individuals. The daily variation of ALP activity observed correlates closely with and is confirmed by the Sigma Histochemical Procedure based on Kaplow' Scoring Method. The coefficient of variation of the ALP method within assay performed on two consecutive days on a pooled sample is 0.75 percent and 1.38 percent, respectively, and between assays 4.82 percent.

UI MeSH Term Description Entries
D007951 Leukemia, Myeloid Form of leukemia characterized by an uncontrolled proliferation of the myeloid lineage and their precursors (MYELOID PROGENITOR CELLS) in the bone marrow and other sites. Granulocytic Leukemia,Leukemia, Granulocytic,Leukemia, Myelocytic,Leukemia, Myelogenous,Myelocytic Leukemia,Myelogenous Leukemia,Myeloid Leukemia,Leukemia, Monocytic, Chronic,Monocytic Leukemia, Chronic,Chronic Monocytic Leukemia,Chronic Monocytic Leukemias,Granulocytic Leukemias,Leukemia, Chronic Monocytic,Leukemias, Chronic Monocytic,Leukemias, Granulocytic,Leukemias, Myelocytic,Leukemias, Myelogenous,Leukemias, Myeloid,Monocytic Leukemias, Chronic,Myelocytic Leukemias,Myelogenous Leukemias,Myeloid Leukemias
D007955 Leukemoid Reaction A peripheral blood picture resembling that of leukemia or indistinguishable from it on the basis of morphologic appearance alone. (Dorland, 27th ed) Leukemoid Reactions,Reaction, Leukemoid,Reactions, Leukemoid
D008297 Male Males
D002469 Cell Separation Techniques for separating distinct populations of cells. Cell Isolation,Cell Segregation,Isolation, Cell,Cell Isolations,Cell Segregations,Cell Separations,Isolations, Cell,Segregation, Cell,Segregations, Cell,Separation, Cell,Separations, Cell
D002498 Centrifugation Process of using a rotating machine to generate centrifugal force to separate substances of different densities, remove moisture, or simulate gravitational effects. It employs a large motor-driven apparatus with a long arm, at the end of which human and animal subjects, biological specimens, or equipment can be revolved and rotated at various speeds to study gravitational effects. (From Websters, 10th ed; McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
D004796 Clinical Enzyme Tests Analyses for a specific enzyme activity, or of the level of a specific enzyme that is used to assess health and disease risk, for early detection of disease or disease prediction, diagnosis, and change in disease status. Enzyme Tests, Clinical,Clinical Enzyme Test,Enzyme Test, Clinical,Test, Clinical Enzyme,Tests, Clinical Enzyme
D006098 Granulocytes Leukocytes with abundant granules in the cytoplasm. They are divided into three groups according to the staining properties of the granules: neutrophilic, eosinophilic, and basophilic. Mature granulocytes are the NEUTROPHILS; EOSINOPHILS; and BASOPHILS. Granulocyte
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000328 Adult A person having attained full growth or maturity. Adults are of 19 through 44 years of age. For a person between 19 and 24 years of age, YOUNG ADULT is available. Adults
D000469 Alkaline Phosphatase An enzyme that catalyzes the conversion of an orthophosphoric monoester and water to an alcohol and orthophosphate. EC 3.1.3.1.

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