cPLA2 Assay Kit

cPLA₂ Assay Kit

Cayman Chemical Item Number 765021

PLA₂ Type IV Assay Kit; Calcium-dependent Cytosolic Phospholipase A₂ Assay Kit

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Description

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'-dithiobis(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.

1 Dennis, E.A. Phospholipases. Enzymes XVI 307-353 (1983).

2 Dennis, E.A. Diversity of group types, regulation, and function of phospholipase A₂. J Biol Chem 269 13057-13060 (1994).

3 Reynolds, L.J., Hughes, L.L., Yu, L., et al. 1-Hexadecyl-2-arachidonoylthio-2-deoxy-sn-glycero-3-phosphorylcholine as a substrate for the microtiterplate assay of human cytosolic phospholipase A₂. Anal Biochem 217 25-32 (1994).

Synonyms

PLA2 Type IV Assay Kit
Calcium-dependent Cytosolic Phospholipase A2 Assay Kit

Stability

1 year

Storage

-20°C

Shipping

Wet ice in continental US; may vary elsewhere

Background Reading

Reynolds, L.J., Hughes, L.L., Yu, L., et al. 1-Hexadecyl-2-arachidonoylthio-2-deoxy-sn-glycero-3-phosphorylcholine as a substrate for the microtiterplate assay of human cytosolic phospholipase A₂. Anal Biochem 217 25-32 (1994).