Isoelectric focusing of human neutrophil alkaline phosphatase isoenzymes in agarose gel.
An isoelectric focusing technique for human neutrophil alkaline phosphatase isoenzymes is described. After sonication with Zwittergent 3-12, butanol extraction and ultracentrifugation, dialysis of cytosols precedes focusing. Focusing patterns show a heterogeneity with two enzymatic activity areas: a main component at pI 6.7-6.8 with a minor component at pI 4.8-5.0 which is difficult to visualize due to its sensitivity to experimental conditions. The addition of 3 mmol/l ZnCl2 to the agarose gel improved the staining of focused bands and in particular the anodic component.
Brisson-Lougarre
A
A
Service Univesitaire d'Hématologie, Faculté de Médecine de Toulouse Rangueil, France.
Vergnes
H
H
Grozdea
J
J
Bierme
R
R
D000469:Alkaline Phosphatase / Q000097:blood*; D002712:Chlorides; D003600:Cytosol / Q000201:enzymology; D003902:Detergents; D003956:Dialysis; D004587:Electrophoresis, Agar Gel / Q000379:methods; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing / Q000379:methods; D007527:Isoenzymes / Q000097:blood; D008099:Liver / Q000201:enzymology; D009504:Neutrophils / Q000201:enzymology*; D011247:Pregnancy; D012016:Reference Values; D015032:Zinc; D017967:Zinc Compounds
D002712:Chlorides; D003902:Detergents; D007527:Isoenzymes; D017967:Zinc Compounds; C016837:zinc chloride; D000469:Alkaline Phosphatase; D015032:Zinc