| In this experiment, the following conditions were applied: Total protein load/well: 30 µg Blocking: 1 h, 5% non-fat milk Primary antibody: 1:1000 O/N 4°C with TBS-T or TBS buffers Secondary antibody: Goat anti-rabbit, 1:25 000 1hr RT with TBS (AS09 602, Agrisera) Detection: Agrisera ECLSuperBright, 5 min As seen in the image to the righ, using TBS buffer without Tween 20 resulted in the detection of the target protein in the overexpressing line and the wild type. |
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