Ubiquilin-2, Proteintech, 23449-1-AP

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 23449-1-AP (antibody dilution 1:1000) produced a strong 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. A band of weaker intensity was detected in both WT and KO cell lysate just below the WT specific band.

IP (2 µg antibody) yielded a distinct band of similar size (approximately 65 kDa), with higher intensity than the corresponding band in the input extract (4% of starting material). Only a faint band was observed in the unbound fraction (4% of total material), indicating complete immunodepletion of the antigen.

In IF (1:300 antibody dilution), intense staining was observed in WT cells with no background staining in KO cells

We highly encourage other scientists to share their observations.