PPP2R5D, Abcam, ab188323

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit delta isoform 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 Serine/threonine-protein phosphatase 2A 56 kDa regulatory subunit delta isoform by PPP2R5D.

The collection of antibodies was tested using HAP1 WT and PPP2R5D KO cells in the applications WB (WT and KO cell lysates/30 µg protein), IP (WT cell lysate) and IF (WT and KO cells) according to YCharOS standard protocols (see reference).

In WB (1:10000 ab dilution) ab188323 detected a band of appr. 65 kDa in WT cell lysate with no corresponding band observed in KO cell lysate. The observed molecular weight of PPP2R5D varies according to the lysis buffer used. HAP1 WT and PPP2R5D KO lysates were prepared in RIPA or IP lysis buffer and the molecular weight of PPP2R5D was assessed by WB using ab188323 at 1:10000 dilution. PPP2R5D ran primarily at ~65 kDa or ~75 kDa when lysates were prepared with RIPA or IP lysis buffer, respectively. A band was also detected following IP (2 μg Ab used). This band was markedly more intense than the corresponding band in input extract (4% of starting material). Band was also detected in immunodepleted extract (4% of total unbound fraction), showing that the antibody does not completely deplete the antigen from the cell lysate. In IF (1:200 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.