A microtiter plate assay for N-acetyl-β-d-glucosaminidase using a fluorogenic substrate

✍ Scribed by S. Linko-Löppönen; M. Mäkinen

• Publisher: Elsevier Science
• Year: 1985
• Tongue: English
• Weight: 295 KB
• Volume: 148
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(85)90626-8

✦ Synopsis

We describe a simple endpoint method for the determination ofN-acetyl-@ghtcosaminidase (NAGase; EC 3.2.1.30). NAGase uses a fluorogenic substrate, 4-methylumbelliferyl-IV-acetyl-@tx-ghtcosaminide, at pH 4.6, liberating the fluorescent 4-methylumbelliferone. The method is reproducible and fast both at room temperature and at 37°C. The procedure developed can be used, e.g., in the diagnosis of bovine subclinical mastitis, where elevated NAGase activities are found in raw milk samples. The assay procedure has a high capacity and high sensitivity and several hundred milk samples can be screened per hour using 96-well microtiter plates and an automated fluorescence reader. In addition to its use in mastitis diagnosis, the assay can be used in the diagnosis of some diseases of human origin. o 1985 Academic PI-M, IIIC.