A study of alkaline phosphatase isoenzyme electrophoresis on cellulose acetate compared with agar, agarose and acrylamide in the presence or absence of triton X-100

1979 ◽  
Vol 91 (2) ◽  
pp. 147-152 ◽  
Author(s):  
H.W. Laan ◽  
D. Diaz ◽  
M. Szakaly
Keyword(s):  
Alkaline Phosphatase ◽  
Cellulose Acetate ◽  
Alkaline Phosphatase Isoenzyme ◽  
Triton X

Quantitative alkaline phosphatase isoenzyme determination by electrophoresis on cellulose acetate membranes.

1977 ◽  
Vol 23 (1) ◽  
pp. 28-34 ◽  
Author(s):  
W H Siede ◽  
U B Seiffert
Keyword(s):  
Alkaline Phosphatase ◽  
Cellulose Acetate ◽  
Clinical Laboratory ◽  
Kinetic Assay ◽  
Specific Staining ◽  
Cellulose Acetate Membranes ◽  
Alkaline Phosphatase Isoenzymes ◽  
Elution Method ◽  
Routine Clinical Laboratory ◽  
Alkaline Phosphatase Isoenzyme

Abstract We present a new method for quantitative determination of alkaline phosphatase isoenzymes. This method consists of electrophoretic separation on cellulose acetate membranes, special fixation technique to avoid elution and diffusion of enzyme protein during incubation, specific staining, and quantitative evaluation by densitometric measurement. We highly recommend the precedure for routine clinical laboratory use. In all normal individuals we observe two isoenzymes of hepatic origin and one isoenzyme each of osseous, intestinal, and biliary origin. Quantitative normal values are presented. Precision of the method is calculated, the CV being less than 10%. The exactness of densitometric quantification is proved by comparison with kinetic assay of alkaline phosphatase isoenzymes by use of an elution method. Clinical implications of alkaline phosphatase isoenzymograms are reported and discussed in detail.

The analysis of alkaline phosphatase isoenzyme using 4-methylumbelliferyl phosphate as substrate on a cellulose acetate membrane

1979 ◽  
Vol 91 (3) ◽  
pp. 273-276 ◽  
Author(s):  
Fukuko Watanabe ◽  
Megumi Takano ◽  
Fumiko Tanaka ◽  
Nobuyuki Amino ◽  
Chozo Hayashi ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Cellulose Acetate ◽  
Cellulose Acetate Membrane ◽  
Alkaline Phosphatase Isoenzyme

scholarly journals Reaction Rate Retardation as a Method for Serum Alkaline Phosphatase Isoenzyme Measurement

1980 ◽  
Vol 17 (4) ◽  
pp. 192-198 ◽  
Author(s):  
Pamela B Brown ◽  
K O Lewis
Keyword(s):  
Alkaline Phosphatase ◽  
Reaction Rate ◽  
Serum Alkaline Phosphatase ◽  
Enzyme Reaction ◽  
Intestinal Alkaline Phosphatase ◽  
Wide Range ◽  
Alkaline Phosphatase Isoenzymes ◽  
Sensitivity And Selectivity ◽  
Rate Retardation ◽  
Alkaline Phosphatase Isoenzyme

A method for serum alkaline phosphatase isoenzymes using an enzyme reaction rate analyser is described. The complete urea-induced degradation of enzyme activity is monitored, from which individual isoenzyme activities are obtained by calculating the constituent exponential components of the degradation curve. Activities have been measured with adequate sensitivity and selectivity for up to four isoenzyme components in normal and in pathological sera. The identity of each isoenzyme present is assigned from its characteristic degradation half-life, and by this method bone and liver alkaline phosphatase are clearly distinguished and quantitated, and a composite value for placental-intestinal alkaline phosphatase activity is obtained. The approach promises to be applicable to a wide range of isoenzymes, and in analogy with ‘reaction rate’ the term ‘reaction rate retardation’ is suggested for the procedure.

Detection of Regan Variant Type of Alkaline Phosphatase Isoenzyme in Liver Tissue of Indian Childhood Cirrhosis

Hepatology ◽  
2007 ◽  
Vol 3 (4) ◽  
pp. 572-576
Author(s):  
S. R. Parekh ◽  
B. D. Patel ◽  
S. R. Damle ◽  
G. M. Thanki ◽  
D. Khutti
Keyword(s):  
Alkaline Phosphatase ◽  
Liver Tissue ◽  
Indian Childhood Cirrhosis ◽  
Variant Type ◽  
Indian Childhood ◽  
Alkaline Phosphatase Isoenzyme

INACTIVATION OF RENIN IN A MIXED MITOCHONDRIAL-LYSOSOMAL FRACTION OF POST-PARTUM UTERUS

1979 ◽  
Vol 92 (4) ◽  
pp. 731-745
Author(s):  
Jørgen Jørgensen
Keyword(s):  
Alkaline Phosphatase ◽  
Post Partum ◽  
Similar Rate ◽  
Freezing And Thawing ◽  
Lysosomal Fraction ◽  
Triton X

ABSTRACT The marked renin inactivation seen during in vitro incubation of post-partum uterine slices which mimics the in vivo condition, is not accompanied by a similar general proteolysis. The inactivating mechanism is so far non-specific with respect to organ or species as added hog renal renin is inactivated at a similar rate as endogenous renin. Endogenous alkaline phosphatase is not significantly inactivated and added alkaline phosphatase is completely stable. A marked inactivation of endogenous renin also takes place during incubation of a mixed mitochondrial-lysosomal suspension prepared from post-partum uterus. The process is more pronounced at pH 7.4 than at 6.8. Freezing and thawing and addition of Triton X-100 prior to incubation inhibits the inactivation. ATP and α-ketoglutarate slightly stimulates the process while CoA and chloroquine have no effect. Both iodoacetate and phenylmethylsulphonylfluoride inhibit the inactivation, suggesting that more than one enzyme is involved in the inactivation.

Incorporation of human liver and placental alkaline phosphatases into liposomes and membranes is via phosphatidylinositol

1990 ◽  
Vol 68 (9) ◽  
pp. 1112-1118 ◽  
Author(s):  
Lee Kihn ◽  
Dorothy Rutkowski ◽  
Robert A. Stinson
Keyword(s):  
Alkaline Phosphatase ◽  
Phospholipase C ◽  
Human Liver ◽  
Red Cell ◽  
Osteosarcoma Cells ◽  
Alkaline Phosphatases ◽  
Membrane Anchor ◽  
Gradient Gel Electrophoresis ◽  
Triton X ◽  
Phosphatidylinositol Phospholipase C

As assessed by incorporation into liposomes and by adsorption to octyl-Sepharose, the integrity of the membrane anchor for the purified tetrameric forms of alkaline phosphatase from human liver and placenta was intact. Any treatment that resulted in a dimeric enzyme precluded incorporation and adsorption. An intact anchor also allowed incorporation into red cell ghosts. The addition of hydrophobic proteins inhibited incorporation into liposomes to varying degrees. Alkaline phosphatase was 100% releasable from liposomes and red cell ghosts by a phospholipase C specific for phosphatidylinositol. There was no appreciable difference in the rates of release of placental and liver alkaline phosphatases, although both were approximately 250 × slower in liposomes and 100 × slower in red cell ghosts than the enzyme's release from a suspension of cultured osteosarcoma cells. Both enzymes were released by phosphatidylinositol phospholipase C as dimers and would not reincorporate or adsorb to octyl-Sepharose. However, the enzyme incorporated, resolubilized by Triton X-100, and cleansed of the detergent by butanol treatment was tetrameric by gradient gel electrophoresis, was hydrophobic, and could reincorporate into fresh liposomes. A monoclonal antibody to liver alkaline phosphatase inhibited the enzyme's incorporation into liposomes, and abolished its release from liposomes and its conversion to dimers by phosphatidylinositol phospholipase C.Key words: alkaline phosphatase, liposome, phosphatidylinositol, membrane anchor.

Alkaline phosphatase isoenzymes.

1982 ◽  
Vol 28 (10) ◽  
pp. 2007-2016 ◽  
Author(s):  
D W Moss
Keyword(s):  
Alkaline Phosphatase ◽  
Quantitative Analysis ◽  
Posttranslational Modifications ◽  
Quantitative Methods ◽  
Molecular Forms ◽  
Future Research ◽  
Alkaline Phosphatases ◽  
Isoenzyme Analysis ◽  
Alkaline Phosphatase Isoenzymes ◽  
Alkaline Phosphatase Isoenzyme

Abstract The human alkaline phosphatases constitute a system of multiple molecular forms of enzymes in which heterogeneity is partly due to genetic factors and partly to posttranslational modifications. Recognition of the nature and occurrence of these multiple forms has made a significant contribution both to the understanding of changes in alkaline phosphatase values for serum in disease and to the use of alkaline phosphatase measurements in diagnosis. Many of the diagnostic advantages of alkaline phosphatase isoenzyme analysis can be obtained with the aid of qualitative methods such as zone electrophoresis. However, quantitative methods are needed to take full advantage of the potential benefits of isoenzyme analysis. Selective inactivation methods can be applied successfully to the quantitative analysis of bone and liver alkaline phosphatases in serum. However, the aim of future research should be to remove the limitations at present imposed on quantitative analysis by the close similarities of bone and liver alkaline phosphatases.

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