TBS or TBS-T, how using respective buffer for primary antibody incubation may change the outcome

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. 



 

TBS or TBS-T buffers for antibody incubation, how can they change the outcome?

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