Cry1JP578V competition binding assays in SBL. (A) Binding of Alexa-labeled Cry1JP578V to SBL BBMV illustrated by homologous competition. BBMVs (30 µg) were incubated with Alexa-Cry1JP578V (2.5 nM) in the absence (“Total”) and presence of unlabeled Cry1JP578V protein. The graphs plot the average densitometry values measured from images taken of in-gel fluorescence (see Section 5.3) for each binding reaction and normalized to the values measured in the absence of competitor, i.e., “% Total Binding”. (B) Heterologous competition between Alexa-labeled Cry1JP578V and representatives of trait proteins with SBL BBMV show “% Total Binding” of Alexa-Cry1JP578V (2.5 nM) to SBL BBMV (30 μg) in the absence (n = 24) and presence of 1 μM unlabeled Cry1JP578V (n = 24), Cry1Ac, Cry1Fa, Cry2A.127, and Vip3Ab (n = 6). (C) Reciprocal competition (n = 6) for 50 nM Alexa Cry1Ac and 5 μg SBL BBMVs; 5 nM Alexa Cry1Fa and 60 μg SBL BBMVs; 20 nM Alexa Cry2A.127 and 30 μg SBL BBMVs; 10 nM Alexa Vip3Ab and 2 μg SBL BBMVs. All the error bars reflect the standard deviation. p-values < 0.01 are indicated by a bracket above the compared columns. Statistical analysis was performed with one-way ANOVA with Tukey’s post hoc test for Panel (B) and with one-way ANOVA with Sidak’s post hoc test for Panel (C).