GPNMB, Thermo Fisher Scientific, MA5-49360

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human GPNMB 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 20 commercial research antibodies against human GPNMB.

The collection of antibodies was tested using U-87 MG WT and GPNMB KD cells in the applications WB (WT and KD cell lysates/60 µg protein), IP (WT cell lysate) and IF (WT and KD cells) according to YCharOS standard protocols (see reference). siRNA technology was used to knockdown (KD) the target gene.

In WB (1:1000 antibody dilution) MA5-49360 detected a double band of appr. 80 and 100 kDa in control lysates not observed in KD lysates. A band was detected following (2 µg Ab used). This band was slightly more intense than the corresponding band observed in input extract (6% of starting material). Band was also detected in immunodepleted extract (6% of total unbound fraction), showing that the antibody does not completely deplete the antigen from the cell lysate. In IF (1:1000 antibody dilution) control cells showed more intense staining intensity, compared to KD cells showing some background.

We highly encourage other scientists to share their observations.