Valosin-containing protein, Aviva Systems Biology, ARP74898

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 ARP74898 (antibody dilution 1:10.000) produced a strong band of approximately 90 kDa in lysate from WT U2OS cells with no corresponding band observed in KD cell lysate. Two weak bands of lower molecular weight were observed in lysates from both WT and KD cells.

IP (using 2 µg antibody) yielded a distinct band at 90 kDa, however band was of comparable intensity to band detected in the input extract (4% of starting material) and in the unbound fraction (4% of total material), indicating that the antigen was not efficiently precipitated.

In IF (1:100 antibody dilution), a clear staining was observed in WT cells with some background staining in KD cells.

We highly encourage other scientists to share their observations.