VPS35, Thermo Fisher Scientific, PA5-21898

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human VPS35 and followed up 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 13 commercial research antibodies against human VPS35.

The collection of antibodies was tested using HAP1 WT and VPS35 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 WB (1:1000 Ab dilution) PA5-21898 detected a band of appr. 75 kDa in WT lysate, with no corresponding band observed in KO lysate. Several weak bands of both high and lower molecular weight were detected in lysates of 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 (2% of starting material). Band was also detected in immunodepleted extract (2% of total unbound fraction), showing that the antibody does not completely deplete the antigen from the cell lysate. In IF (1:100 Ab dilution), comparable staining intensities were observed in both KO and WT cells.

We highly encourage other scientists to share their observations.