Valosin-containing protein, Abcam, ab110308

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Valosin-containing protein (VCP) 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 16 commercial research antibodies against human VCP.

The collection of antibodies was tested using U2OS WT cells and cells with siRNA mediated knock-down of VCP (VCP KD) in the in the applications WB, IP and IF according to YCharOS standard protocols (see reference).

WB was performed using lysates (10 µg protein) from WT and KD cells. IP was performed using WT cell lysates, and immunoprecipitated fraction was analysed by WB using anti-VCP MA3-004 from Thermo Fisher Scientific. IF staining was performed using WT and KD cells.

WB with ab110308 (antibody dilution 1:10.000) produced a single weak band of approximately 50 kDa in lysate from both WT and KD U2OS cells.

Although the antigen was clearly detected in the starting material (4% of total material) and unbound (4% of total material) fractions, no corresponding band was observed in the immunoprecipitated fraction (2 µg antibody used). In addition, a 50 kDa band was detected in the immunoprecipitated fraction, likely due to cross-reactivity of the WB secondary antibody with the Ig heavy chain

In IF (1:1000 antibody dilution), comparable staining intensities were observed in both KD and WT cells.

We highly encourage other scientists to share their observations.