A rapid method for the estimation of beta-galactocerebrosidase, beta-glucocerebrosidase and sphingomyelinase activities in leukocytes.
A method for the assay of leukocyte beta-galactocerebrosidase, beta-glucocerebrosidase and sphingomyelinase activities has been developed, based on the separation of the tritiated sphingolipid substrates from their corresponding radioactive hydrophobic product (ceramide) by thin-layer chromatography on Silica gel H coated microscope slides. For the determination of beta-galactocerebrosidase and beta-glucocerebrosidase activities the silica gel is impregnanted with sodium tetraborate. Each chromatogram is easily divided into two distinct zones and the radioactivity content of each is determined by liquid scintillation counting. The technique described, is rapid, less costly than conventional methods and provides an accurate assessment of sphingolipid hydrolase activity. It is suggested that it should be of considerable value in those areas which require the rapid analysis of large numbers of samples, such as in screening for the sphingolipidoses or for enzyme purification studies.
D002849:Chromatography, Gas; D002855:Chromatography, Thin Layer; D005696:Galactosidases / Q000097:blood*; D005698:Galactosylceramidase / Q000097:blood*; D005959:Glucosidases / Q000097:blood*; D005962:Glucosylceramidase / Q000097:blood*; D006801:Humans; D007700:Kinetics; D007962:Leukocytes / Q000201:enzymology*; D008722:Methods; D010727:Phosphoric Diester Hydrolases / Q000097:blood*; D013108:Sphingomyelin Phosphodiesterase / Q000097:blood*; D013997:Time Factors
D010727:Phosphoric Diester Hydrolases; D013108:Sphingomyelin Phosphodiesterase; D005696:Galactosidases; D005959:Glucosidases; D005962:Glucosylceramidase; D005698:Galactosylceramidase