[Comparison of spectrophotometric and fluorimetric methods of determination of galactocerebroside-beta-D-galactosidase in leukocytes].
A procedure was developed for estimation of galactocerebroside beta-D-galactosidase activity in leukocytes using a new fluorogenic compound 6-hexadecanoylamino-4-methylum-belliferyl-beta-D-galactop yra noside (HMGal) as a substrate. Some patterns of the fluorometric procedure were compared with corresponding parameters of the spectrophotometric method in which a chromogenic substrate HNGal was used. Sensitivity of the fluorometric procedure with HMGal as a substrate was increased 100-fold as compared with the spectrophotometric method. At the same time, the fluorometric procedure enabled to reduce considerably the incubation period and the cell protein content per an assay. High sensitivity and reproducibility of the procedure with HMGal as a substrate allowed to carry out biochemical diagnosis of Krabbe disease in leukocytes.
Gusina
N B
NB
Tsukerman
G L
GL
D005470:Fluorometry; D005696:Galactosidases / Q000097:blood*; D005698:Galactosylceramidase / Q000097:blood*; D006801:Humans; D007962:Leukocytes / Q000201:enzymology*; D013050:Spectrometry, Fluorescence
D005696:Galactosidases; D005698:Galactosylceramidase
Sravnenie spektrofotometricheskogo i fluorimetricheskogo metodov opredeleniia aktivnosti galaktotserebrozid-beta-D-galaktozidazy v leĭkotsitakh.