An improved fluorimetric determination of alpha-mannosidase activity in bovine plasma.
This paper describes a manual fluorimetric method for the assay of acidic alpha-mannosidase activity in bovine plasma. The optimum conditions for the assay of this enzyme were studied. The assay method devised includes the addition of zinc to the substrate, which stimulates activity by approximately twofold, and reduces the optimum substrate concentration. This latter feature affords considerable cost saving in each test. We have also shown that the alpha-mannosidase activity in lithium heparin plasma, EDTA plasma and blood serum is the same whether the plasma/serum is separated from the cells/clot at 6, 20 or 25 hours after sample collection. This has eliminated the previous necessity of having to deliver whole blood samples to the laboratory within 6 hours of collection. Furthermore samples for the supplementary neutrophil assay can now be taken at the same time as those for the plasma test, and both samples forwarded together. The plasma alpha-mannosidase assay is a rapid and reliable screening test for the mannosidosis genotype and for detecting carrier animals. Carrying out this plasma assay in conjunction with the more definitive neutrophil assay provides a reliable method of distinguishing homozygotes and heterozygotes from normal animals.
D000818:Animals; D001681:Biological Assay; D001777:Blood Coagulation; D001800:Blood Specimen Collection; D002417:Cattle / Q000097:blood*; D004492:Edetic Acid / Q000097:blood; D005260:Female; D005470:Fluorometry / Q000379:methods; D006493:Heparin / Q000097:blood; D006863:Hydrogen-Ion Concentration; D008297:Male; D008361:Mannosidases / Q000097:blood*; D015032:Zinc / Q000494:pharmacology
D006493:Heparin; D004492:Edetic Acid; D008361:Mannosidases; D015032:Zinc