Dnmt1, Cell Signaling Technology, 5032

I am sharing a pAbmAbs generated literature review of KO validated antibodies. Here citing Wu et al. Journal for ImmunoTherapy of Cancer, 2021.
Wu et al. validate this antibody for detection of endogenous Dnmt1, by Western blotting of BT-549 and HCC1143, two triple-negative breast cancer cell lines. For both cell lines three KO validations were made with sgDNMT1.
BT-549 sgDNMT1 showed a faint residual band at #3 KO while HCC1143 likely retained partial DNMT1 expression, showed in a strong band at HCC1143#3 KO – either incomplete editing, mixed clone, or partial protein persistence has happened. This doesn’t indicate off-target binding; the same antibody cleanly loses signal in BT-549 KO, showing specificity (Supplementary figure 7B).

Based on the available data, we have assigned a four-star rating because of slight residual signal, but we highly encourage other scientists to share their observations.