cPLA₂ Assay Kit

Phospholipase A₂ catalyzes the hydrolysis of fatty acids at the sn-2 position of glycerophospholipids, yielding a free fatty acid and a lysophospholipid as products.¹ The release of arachidonic acid from membrane phospholipids by these enzymes is believed to be the key step in the biosynthesis of eicosanoids.² There are primarily three different kinds of phospholipase A₂. They are secretory (sPLA₂), calcium-dependent cytosolic (cPLA₂), and calcium-independent cytosolic (iPLA₂) phospholipase A₂. Of these three different types of enzymes, only the cPLA₂ exhibits specificity towards arachidonic acid whereas all others can hydrolyze any fatty acid at the sn-2 position. Arachidonoyl Thio-PC is a substrate for cPLA₂ by virtue of the presence of arachidonic acid at the sn-2 position of the glycerophospholipid.³ Hydrolysis of the arachidonoyl thioester bond at the sn-2 position by PLA₂ releases a free thiol which can be detected by DTNB (5,5'-dithio-bis-(2-nitrobenzoic acid)). This assay can be used to determine the activity of cPLA₂ in purified preparations, cell cultures, or tissue homogenates that are known to contain only cPLA₂. Use of this assay with preparations containing more than one type of PLA₂ will result in the measurement of total PLA₂ activity rather than cPLA₂ alone. Isozyme-specific cPLA₂ activity can be measured by excluding sPLA₂ or inhibiting iPLA₂ activities in the assay. Each kit contains cPLA₂ assay buffer, DTNB/EGTA, Arachidonoyl Thio-PC (substrate), bee venom PLA₂ (control), bromoenol lactone solution, a 96 well plate, and complete instructions.

Needed but not supplied: Please download the kit booklet to verify if - (Milli-Q or equivalent) or any other components are needed for this assay.

WARNING This product is not for human or veterinary use.