Fig. 4.

The effect of 0, 6, 20, 40, 60, 200, and 600 μM cocaine on the non-reducing SDS-PAGE banding pattern for the control intact h2E2 mAb and the deglycosylated intact h2E2 mAb. 10 μL, containing 2 μg of each sample was loaded in each well of a 7% acrylamide non-reducing SDS-PAGE gel, and stained with Coomassie blue. The concentration of cocaine added to the samples are indicated above each well. The red dashed lines indicate the electrophoretic migration positions of the totally denatured proteins (proteins boiled for 5 min in the SDS sample buffer without a reductant). No bands were visible in the bottom sections of these two gels, which were cropped to make a more compact figure. The vertical black line was superimposed on the image of the two aligned gels shown in Panels A and B, and each Panel image was from a single gel, with the samples loaded in the order shown. Panel A (top) shows the banding pattern gel results when the samples were run less than 5 min after dilution into the non-reducing SDS-PAGE sample buffer (the same sample treatment used in Fig. 1, Fig. 2, Fig. 3), while Panel B (bottom) shows the banding pattern of aliquots of the same samples shown in Panel A, after incubation of those samples in the sample buffer for 24 h at 22 °C. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)