Table 17.3 Fluorescence-based phospholipase assays.

Phospholipase * Probes Assay Principle Detection Method
A1 A10070, E10219, E10221 Intramolecular self-quenching Fluorescence increase at ~530 nm
A1, A2 B7701 Intramolecular self-quenching Fluorescence increase at ~515 nm ref
A1, A2 B3781, B3782 Intramolecular excimer formation Emission ratio 380/470 nm ref
A1, A2 A3880  Free fatty acid sensor Emission ratio 432/505 nm ref
A2 A10072, E10217, E10218 Intramolecular fluorescence resonance energy transfer (FRET) Fluorescence increase at ~515 nm or increase in emission ratio at 515/575 nm
A2 D23739 Intramolecular self-quenching Fluorescence increase at ~515 nm ref
A2 N3786, N3787 Intermolecular self-quenching Fluorescence increase at ~530 nm ref
A2 H361, H3809 Intermolecular excimer formation Emission ratio 380/470 nm ref
A2 D3803 Release of a fluorescent fatty acid TLC or fluorescence image scanner ref
A2, C, D D3771 Formation of a fluorescent O-alkylglycerol derivative TLC or HPLC ref
A2, C, D H361 Quenching by a disulfide-polymerized lipid matrix Fluorescence increase at ~380 nm ref
C A12218 Peroxidase-linked detection of phosphocholine Conversion of the nonfluorescent Amplex Red reagent to fluorescent resorufin ref
C E10215, E10216 Release of dye-labeled diacylglycerol Fluorescence increase at 516 nm, with potential interference from phospholipase A2 and phospholipase D activity
D A12219 Peroxidase-linked detection of choline Conversion of the nonfluorescent Amplex Red reagent to fluorescent resorufin ref
PAP † D3805 Release of dye-labeled diacylglycerol HPLC ref
* Phospholipase specificity: A1, A2, C or D (see Probes for Lipid Metabolism and Signaling—Section 17.4 for cleavage specificities). † PAP = phosphatidic acid phosphohydrolase.