Parkin, Thermo Fisher Scientific, 702785

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 parkin 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 16 commercial research antibodies against human E3 ubiquitin-protein ligase parkin encoded by PRKN.

The collection of antibodies was tested using SH-SY5Y WT and PRKN KO cells in the applications WB (WT and KO cell lysates/50 µg protein), IP (WT cell lysate) and IF (WT and KO cells) according to YCharOS standard protocols (see reference). Furthermore, this antibody was tested in HEK293T, SH-SY5Y, HAP1, HeLa, U2OS, MDA-MB231, A549 and HCT116 in the application WB (KO or WT cell lysate/100 µg protein).

In WB (1:200 ab dilution), 702785 detected a band of appr. 50 kDa in WT cell lysate, with no corresponding band observed in KO cell lysate. A band of higher molecular weight was observed in both genotypes. In Western blotting (1:200 antibody dilution), a band at approximately 50 kDa was detected in WT lysates from SH-SY5Y, HAP1, and HEK293T cells. A band was also detected following IP (1 µg Ab used), this band was markedly more intense than corresponding band in input extract (10% of starting material), no band was detected in immunodepleted extract (10% of total unbound fraction), showing that the antibody does deplete the antigen from cell lysate. In IF (1:500 ab dilution), comparable staining intensity were observed in both WT and KO cells.