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β‐Galactosidase assay using capillary electrophoresis laser‐induced fluorescence detection and resorufin‐β‐ D ‐galactopyranoside as substrate
Author(s) -
Eggertson Michael J.,
Craig Douglas B.
Publication year - 1999
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/(sici)1099-0801(199912)13:8<516::aid-bmc918>3.0.co;2-u
Subject(s) - chemistry , capillary electrophoresis , chromatography , substrate (aquarium) , fluorescence , detection limit , enzyme , laser induced fluorescence , electrophoresis , biochemistry , oceanography , physics , quantum mechanics , geology
β‐Galactosidase was incubated for 60 min with the fluorogenic substrate resorufin‐β‐ D ‐galactopyranoside, which is converted by the action of the enzyme into resorufin and galactose. A 160 pL aliquot of reaction mixture was analyzed by capillary electrophoresis utilizing laser‐induced fluorescence detection. Based on the detection of the resorufin formed, the limit of detection of β‐galactosidase was 1.5 × 10 −15 M or 900 molecules of enzyme in a 1 µL sample. Copyright © 1999 John Wiley & Sons, Ltd.