Table 17.3 Fluorescence-based phospholipase assays.

Phospholipase * Probes Assay Principle Detection Method
A1A10070, E10219, E10221Intramolecular self-quenchingFluorescence increase at ~530 nm
A1, A2B7701Intramolecular self-quenchingFluorescence increase at ~515 nm ref
A1, A2B3781, B3782Intramolecular excimer formationEmission ratio 380/470 nm ref
A1, A2A3880 Free fatty acid sensorEmission ratio 432/505 nm ref
A2A10072, E10217, E10218Intramolecular fluorescence resonance energy transfer (FRET)Fluorescence increase at ~515 nm or increase in emission ratio at 515/575 nm
A2D23739Intramolecular self-quenchingFluorescence increase at ~515 nm ref
A2N3786, N3787Intermolecular self-quenchingFluorescence increase at ~530 nm ref
A2H361, H3809Intermolecular excimer formationEmission ratio 380/470 nm ref
A2D3803Release of a fluorescent fatty acidTLC or fluorescence image scanner ref
A2, C, DD3771Formation of a fluorescent O-alkylglycerol derivativeTLC or HPLC ref
A2, C, DH361Quenching by a disulfide-polymerized lipid matrixFluorescence increase at ~380 nm ref
CA12218Peroxidase-linked detection of phosphocholineConversion of the nonfluorescent Amplex Red reagent to fluorescent resorufin ref
CE10215, E10216Release of dye-labeled diacylglycerolFluorescence increase at 516 nm, with potential interference from phospholipase A2 and phospholipase D activity
DA12219Peroxidase-linked detection of cholineConversion of the nonfluorescent Amplex Red reagent to fluorescent resorufin ref
PAP †D3805Release of dye-labeled diacylglycerolHPLC 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.

For Research Use Only. Not for use in diagnostic procedures.