Improved agarose electrophoretic method for separating alkaline phosphatase isoenzymes in serum.
A modified agarose electrophoretic system for the separation of alkaline phosphatase (ALP, EC 3.1.3.1) isoenzymes is described. Bone, liver, high-molecular-mass, and intestinal ALP are separated with high reproducibility. The sensitivity of the agarose system is superior to cellulose acetate in detecting high-Mr ALP. Correlation is good between bone ALP fractions scanned before and after treatment with neuraminidase. Immunoglobulin-bound ALPs, the ALP-lipoprotein-X complex, and the additional ALP fraction observed in transient hyperphosphatasemia in children are detected by their peculiar electrophoretic mobility in the proposed system. Approximately 25% of the samples contained an additional fraction of intestinal-type ALP, as evidenced by neuraminidase treatment and use of polyclonal and monoclonal antibodies. Because the electrophoretic mobilities of this "intestinal variant" and of some immunoglobulin-bound ALP fractions are identical to those of bone and intestinal ALP, respectively, treatment of the samples with a polyclonal antibody that reacts with intestinal ALP is advised.
D000469:Alkaline Phosphatase / Q000097:blood* / Q000235:genetics; D001842:Bone and Bones / Q000201:enzymology; D004586:Electrophoresis; D004587:Electrophoresis, Agar Gel; D004588:Electrophoresis, Cellulose Acetate; D005260:Female; D014644:Genetic Variation; D006509:Hepatitis B / Q000201:enzymology; D006801:Humans; D007163:Immunosorbent Techniques; D007223:Infant; D007422:Intestines / Q000201:enzymology; D007527:Isoenzymes / Q000097:blood*; D008099:Liver / Q000201:enzymology; D008970:Molecular Weight; D009439:Neuraminidase / Q000494:pharmacology; D010710:Phosphates / Q000097:blood; D010920:Placenta / Q000201:enzymology
D007527:Isoenzymes; D010710:Phosphates; D000469:Alkaline Phosphatase; D009439:Neuraminidase