Protein Extraction and Electrophoresis of A. ceylanicum ESPs

Approximately 3 μg of A. ceylanicum adult ESPs was mixed with 2× concentrated Laemmli-SDS buffer (Bio-Rad) with 5% 2-Mercaptoethanol (Sigma-Aldrich). The sample was heated at 95 °C for 5 min and 50 µL of protein sample was loaded into the wells of 12% Mini-PROTEAN TGX gel (Bio-Rad). The precision protein standard (Bio-Rad) was loaded into the first well at 10 µL without dilution with an SDS sample buffer. Electrophoresis was run for 50 min at 150 V using 1× Tris/Glycine buffer (Bio-Rad), [6 (link)]. The gel was fixed in (40% ethanol and 10% acetic acid), washed with distilled water, and stained with QC Colloidal Coomassie Stain (Bio-Rad). Gels were imaged in Azure Imaging Systems (C600, Azure Biosystems, Dublin, CA, USA) under LED visible fluorescence detection mode (data not shown).