Sequestosome-1, Abcam, ab109012

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 (1:10000 ab dilution) ab109012 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 was markedly more intense compared to band in input extract (10% of starting material) and no band was detected in immunodepleted extract (10% of total unbound fraction), showing that the antibody does completely deplete the antigen from cell lysate. In IF (1:500 ab dilution), a more intense staining was observed in WT cells compared to KO cells showing some background.

I am sharing a pAbmAbs generated literature review of KO validated antibodies. Here citing Yu et al. Frontiers in Oncology 2021.
Yu et al. validate this antibody for detection of human TPC-1 cell line by Western blotting of WT and p62-KO TPC-1 cells. No band was observed in p62-KO (figure 1d).
Based on the available data, we have assigned a four-star rating because of missing image of full blot, but we highly encourage other scientists to share their observations.