Cell Signaling Technology
Phospho-Smad3 (Ser423/425) (C25A9) Rabbit mAb
9520
C25A9
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We used this antibody to detect phospho SMAD3 in primary mouse naive CD4+ T-cells treated with different doses of SMAD3 inhibitor. The antibody had numerous non-specific bands, especially above 100 kDa, many of which were brighter than the bands at the predicted MW.
Western Blot
Primary mouse CD4+ T cells treated with varying doses of SMAD3 inhibitor
1:1000, overnight, at 4 C
5% BSA
1:2000 anti-Rabbit HRP-linked
N/A
Chemiluminescence
We found that the antibody was able to detect phospho-SMAD3, but the band was quite faint compared to many non-specific bands.This made quantification very challenging and we did not feel confident in these results.
None
Too many non-specific bands in our detection, made quantification difficult
There are other antibodies for detection of Phospho-SMAD3 that produce a much cleaner image for western blot quantification.