Ubiquilin-2, Thermo Fisher Scientific, 37-7700

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Ubiquilin-2 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 10 commercial research antibodies against human Ubiquilin-2 encoded by UBQLN2.

The collection of antibodies was tested using HAP1 WT and UBQLN2 KO cells in the applications WB, IP and IF according to YCharOS standard protocols (see reference).

WB was performed using lysates (30 µg protein) from WT and KO cells. IP was performed using WT cell lysates, and immunoprecipitated fraction was analysed by WB using anti-Ubiquilin-2 85509 (1:1000) from Cell Signaling Technology. IF staining was performed using WT and KO cells.

WB with 37-7700 (antibody dilution 1:2000) produced a weak band between the 48 kDa and 75 kDa markers in lysates from HAP1 WT cells, with no corresponding band observed in KO lysate. This matches the predicted size of 65 kDa for Ubiquilin-2. However, a band of similar intensity was detected in both WT and KO cell lysate just below the WT specific band, in addition to a faint band just above the specific band in cells of both genotypes.

IP (2 µg antibody) yielded a distinct band of approximately 65 kDa of comparable intensity as the corresponding band in the input extract (4% of starting material) and in the unbound fraction (4% of total material), indicating poor immunodepletion of the antigen.

In IF (1:500 antibody dilution), staining of comparable intensities was observed in WT and KO cells.

We highly encourage other scientists to share their observations.