Abstract
The prophenoloxidase activating system (ProPO-AS) is an integral part of the constitutive in- nate immune response in insects, the products of which are commonly assayed to assess an individual's ability to respond to immune challenges. However, there is considerable variation in the methodologies associated with these assays, and as such, it is not always clear how to interpret results. We have optimised assays for measuring phenoloxidase in its active (PO) and zymogen (ProPO) forms in the honey bee, Apis mellifera. Contrary to results for other insects, we found that the activator α-chymotrypsin, when used at a low concentration (0.5 mg mL −1 ), combined with a minimal activation time (5 min), provided optimal conditions for assaying ProPO. In addition, a saturated L-dopa solution was required for assaying both PO and ProPO. The results highlight the importance of defining the working parameters of each assay to be species-specific. honey bee / Apis mellifera / phenoloxidase / innate immunity / activator
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