Rabbit anti ctb polyclonal primary antibody

Total plant protein from the greenhouse grown young leaves or the powdered lyophilized plant cells of transplastomic lettuce expressing CTB-VP1n and CTB-VP1co, and wild-type were extracted in the protein extraction buffer (100 mM NaCl, 10 mM EDTA, 200 mM Tris-Cl pH 8.0, 0.05% (v/v) Tween-20, 0.1% SDS, 14 mM β-ME, 400 mM sucrose, 2 mM PMSF, and proteinase inhibitor cocktail). Total protein extracts were analyzed by 10% SDS-PAGE. Western blot analysis was performed by rabbit anti-CTB polyclonal primary antibody (GenWay Biotech Inc, San Diego, CA) in a dilution of 1:10,000 followed by detection with a goat anti-rabbit IgG-HRP secondary antibody (1: 4,000 dilution; SouthernBiotech, Birmingham, AL) and developed using the ECL chemiluminescence (GE healthcare, UK). CTB (Cat# GWB-7B96E4, GenWay Biotech Inc, San Diego, CA) was used as positive control and served as standard for quantitative analysis of the blot with Image J software (IJ 1.46r; NIH). For loading controls, protein-blotted membrane was stained with Ponceau S (Sigma, P-3504) prior to immunoprobing with anti-CTB antibody and anti-RbcL antibody (Cat# AS03 037, Agrisera, 1:5,000) was used.