- Enzymatic Assays and Enzyme Histochemistry ofTuta absoluta Feeding on Tomato Leaves.
Enzymatic Assays and Enzyme Histochemistry ofTuta absoluta Feeding on Tomato Leaves.
Bio-protocol (2018-09-05)
Rim Hamza, José P Beltrán, Luis A Cañas
PMID34395792
ABSTRACT
Enzymes play a key role in insect-plant relationships. For a better understanding of these interactions, we analyzed Tuta absoluta digestive enzymes. Here, we describe a detailed protocol for the detection of trypsin and papain-like enzymes in Tuta absoluta larvae by enzyme histochemistry. This assay uses frozen and unfixed samples to avoid the loss of enzymatic activity. We also describe a protocol for the quantification of trypsin and papain-like enzymes in the larvae of Tuta absoluta at different developmental instars.
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
Nα-Benzoyl-L-arginine 4-nitroanilide hydrochloride, ≥99% (TLC), suitable for substrate for trypsin
Sigma-Aldrich
Trypsin from bovine pancreas, TPCK Treated, essentially salt-free, lyophilized powder, ≥10,000 BAEE units/mg protein