VAPB, ABclonal, A5363

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human vesicle-associated membrane protein (VAMP)-associated protein B and C (VAPB) 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 6 commercial research antibodies against human VAPB.

The collection of antibodies was tested using HeLA WT and VAPB KO cells in the applications WB, IP and IF according to YCharOS standard protocols (see reference).

WB was performed using lysates (40 µg protein) from WT and KO cells. IP was performed using WT cell lysates (1 mg), and immunoprecipitated fraction was analysed by WB using anti-VAPB ab315013 (1:1000) from Abcam. IF staining was performed using WT and KO cells.

WB with A5363 (antibody dilution 1:500) produced a strong band between the 25 kDa and 35 kDa markers in lysates from both HeLa WT and KO cells. In addition, several weak bands of higher molecular weight were observed, with some variability between WT and KO 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 IF (1:400 antibody dilution), staining of comparable intensities was observed in WT and KO cells.

We highly encourage other scientists to share their observations.