Sequestosome-1, Abcam, ab56416

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Sequestosome-1 and followed up by extraction and evaluation of the available data to assess its performance according to pAbmAbs rating criteria.

YCharOS is here conducting a large head-to-head study comparing 17 commercial research antibodies against human Sequestosome-1 encoded by SQSTM1.

The collection of antibodies was tested using U2OS WT and SQSTM1 KO cells in the applications WB (WT and KO cell lysates/25 µg protein), IP (WT cell lysate) and IF (WT and KO cells) according to YCharOS standard protocols (see reference).

In WB (1:1000 ab dilution) ab56416 detected a band of appr. 60 kDa in WT cell lysate with no corresponding band observed in KO cell lysate. A band was detected following IP (1 µg Ab used), this band had same intensity as band in input extract (10 % of starting material) and a faint band was observed in immunodepleted extract (10 % of total unbound fraction), showing that the antibody does not efficiently deplete the antigen from the cell lysate. In IF (1:1000 ab dilution), clear staining was observed in WT cells, whereas KO cells showed no staining.

I am sharing a pAbmAbs generated literature review of KO validated antibodies. Here citing Yoon et al. Nature Communications 2021.
Yoon et al. validate this antibody for detection of endogenous human p62 by Immunostaining of p62 knockout HeLa cells (HeLa/p62KO). No staining was observed in control vector while Fig. 1b shows p62 staining in those cells after transfection with FLAG-p62 (WT or mutant) (Figure 1b).
Based on the available data, we have assigned a five-star rating, but we highly encourage other scientists to share their observations.