✦ LIBER ✦

β-Galactosidase assay using capillary electrophoresis laser-induced fluorescence detection and resorufin-β-D-galactopyranoside as substrate

✍ Scribed by Michael J. Eggertson; Douglas B. Craig

Book ID: 101277342
Publisher: John Wiley and Sons
Year: 1999
Tongue: English
Weight: 50 KB
Volume: 13
Category: Article
ISSN: 0269-3879
DOI: 10.1002/(sici)1099-0801(199912)13:8<516::aid-bmc918>3.0.co;2-u

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✦ Synopsis

b-Galactosidase was incubated for 60 min with the fluorogenic substrate resorufin-b-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 b-galactosidase was 1.5 Â 10 À15 M or 900 molecules of enzyme in a 1 mL sample.

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