Neurosecretory protein VGF, Abcam, ab69989

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human neurosecretory protein VGF 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 7 commercial research antibodies against human VGF.
The collection of antibodies was tested using U2OS WT and Vgf KO cells in the applications WB and IP according to YCharOS standard protocols (see reference). WB was performed using lysates (100 µg protein) from WT and KO cells with and without Brefeldin A treatment and cell culture supernatant. IP was performed using WT and KO cell culture supernatant. Western blotting with ab69989 (antibody dilution 1:2000) produced a clear intense band just below the 100 kDa marker in WT cell culture supernatant, with no corresponding signal in KO conditioned media. In addition, several bands of lower molecular weight were observed in conditioned media from both WT and KO cells.

In brefeldin A–treated cells, a double band around the 100 kDa marker was observed, indicating possible accumulation of VGF precursor species of different molecular weights in the secretory pathway. A weak ~100 kDa band was also detected in lysates from untreated WT cells, and in KO lysate from treated and untreated cells. Multiple background bands (intense and weak) were observed in both WT and KO lysates.

Immunoprecipitation (1 µg antibody) resulted in 3 bands, upper band just above the 100 kDa marker. However, the precipitated band displayed a slight shift in apparent molecular weight compared to size of band in input extract, and no increased intensity could be observed. We highly encourage other scientists to share their observations.