A rapid fluorometric assay for the determination of serum phospholipase A2 activity

✍ Scribed by M. Merchant; J. Small; R. Falconi; B.J. Crookshank

• Publisher: Elsevier Science
• Year: 2009
• Tongue: English
• Weight: 241 KB
• Volume: 91
• Category: Article
• ISSN: 0026-265X
• DOI: 10.1016/j.microc.2008.08.006

✦ Synopsis

Phospholipase A 2 (PLA 2 ) is an enzyme that cleaves fatty acids from the sn-2 position of membrane phospholipids. Intracellular PLA 2 has been shown to be important for the generation of arachidonic acid, which leads to the synthesis of a wide range of eicosanoid paracrine hormones. Furthermore, secretory PLA 2 in the serum has been reported to exhibit important immune activities. In this study, BODIPY™ FL C 16 was used as a fluorescent probe to measure PLA 2 activity in the serum of the American alligator. The activity measured was dependent on the amounts of serum and labeled bacteria incubated. The phospholipase activity was also time-dependent, with near peak activities observed within 20-30 min. In addition, the observed activity was titrated away by increasing concentrations of unlabeled bacteria, indicating that the activity observed was specifically due to the presence of PLA 2 . The assay provides rapid results and requires only small amounts of serum.