TMEM175, Abcam, ab300457

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Endosomal/lysosomal proton channel TMEM175 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 6 commercial research antibodies against human Endosomal/lysosomal proton channel TMEM175 encoded by TMEM175. The collection of antibodies was tested using HeLa WT and TMEM175 KO cells in the applications WB (WT and KO cell lysates/20 µg protein), IP (WT cell lysate) and IF (WT and KO cells) according to YCharOS standard protocols (see reference). In addition, the antibody was tested in a broader panel of cell lines, including HeLa WT and TMEM175 KO as well as A172, DM S53, U-87 MG and SH-SY5Y.

In WB (1:1000 ab dilution) ab300457 detected a band of appr. 42 kDa in WT cell lysate with no corresponding band observed in KO cell lysate. Bands of higher molecular weight was observed in both genotypes. Across the different cell types, the strongest band was observed in DMS 53 cells; however, all cell lines showed a band at approximately 42 kDa. A band was detected following IP (2 µg Ab used), this band was markedly more intense compared to bands in input extract (4% of starting material) and immunodepleted extract (4% of total unbound fraction), showing that the antibody does participate most of the antigen; however, not all. In IF (1:250 ab dilution) a more intense staining was observed in some WT cells while many WT and KO cells were comparable.

We highly encourage other scientists to share their observations.