Itch, Abcam, ab280211

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human E3 ubiquitin-protein ligase Itchy homolog 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 9 commercial research antibodies against human E3 ubiquitin-protein ligase Itchy homolog encoded by Itch.

The collection of antibodies was tested using HeLa WT and ITCH KO cells in the applications WB (WT and KO cell lysates/100 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) ab280211 detected a band of appr. 100 kDa in WT cell lysate not observed in KO cell lysate. Two bands of lower molecular weight and with stronger intensity was observed in both genotypes. A band was also detected following IP (2 µg Ab used). This band was more intense than the corresponding band in input extract (10% of starting material). A faint band was detected in immunodepleted extract (10% of total unbound fraction), showing that the antibody does not deplete the antigen from the cell lysate. In IF (1:1000 ab dilution) comparable staining intensities were observed in both KO and WT cells.

We highly encourage other scientists to share their observations.