Ataxin-2, Thermo Fisher Scientific, PA5-54183

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Ataxin-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 five commercial research antibodies against human Ataxin-2 encoded by ATXN2.

The collection of antibodies was tested using HAP1 WT and ATXN2 KO cells in the applications WB (WT and KO cell lysates/50 µg protein), IP (WT cell lysate) and IF (WT and KO cells) according to YCharOS standard protocols (see reference).

Western blot (WB) analysis was performed for both short and long exposure times to evaluate antibody signal strength and specificity. In WB (1:2000 Ab dilution) PA5-54183 detected a band of appr. 140 kDa with no corresponding band in KO for short or long exposure time. Several bands of both high and lower molecular weight were detected in lysates of both genotypes. A band was also detected following IP (1 µg Ab used). This band was not more intense than the corresponding band in input extract (10% of starting material). Band was also detected in immunodepleted extract (10% of total unbound fraction), showing that the antibody does not completely deplete the antigen from the cell lysate. In IF (1:500 Ab dilution), a clear intense staining was observed in WT cells with no background staining in KO cells.

We highly encourage other scientists to share their observations.