, Thermo Fisher Scientific, 14-9711-82

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Ras-related protein Rab5A 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 11 commercial research antibodies against human Ras-related protein Rab5A encoded by RAB5A.

The collection of antibodies was tested using HAP1 WT and RAB5A KO cells in the applications WB (WT and KO cell lysates/30 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) 14-9711-82 detected a faint band of appr. 24 kDa in WT cell lysate with no corresponding band observed in KO cell lysate. A band was detected following IP (2 ug ab used). This band was markedly more intense than the corresponding band in input extract (4% of starting material). A band was also detected in immunodepleted extract (4% of total unbound fraction), showing that the antibody does not completely deplete the antigen from the cell lysate. In IF (1:1000 ab dilution), a more intense staining was observed in WT cells compared to KO cells showing some background.

We highly encourage other scientists to share their observations.