GPNMB, R&D Systems, AF2550

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 (1:500 Antibody dilution) AF2550 detected a double band of appr. 80 and 100 kDa in control lysate not observed in KD lysate. A band was also detected following IP (2 µg Ab used). This band was markedly more intense than the corresponding band 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 compared to KD cells, showing no staining.

We highly encourage other scientists to share their observations.