PARP-1, Cell Signaling Technology, 9532

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Poly (ADP-ribose) polymerase-1 (PARP-1) 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 25 commercial research antibodies against human Poly (ADP-ribose) polymerase-1 (PARP-1).

The collection of antibodies was tested using HAP1 WT and PARP1 KO cells in the applications WB (WT and KO cell lysates/30 µg protein), IP (WT cell lysate), IF (WT and KO cells) and flow cytometry (KO and WT cells) according to YCharOS standard protocols (see reference). This antibody was also tested in HEK293T WT and KO in the application WB.

In WB (1:1000 ab dilution) 9532 detected a band of appr. 120 kDa in WT cell lysate with no corresponding band observed in KO cell lysate. This pattern was consistent in both cell lines. A band was detected following IP (2 µg Ab used), this band was more intense compared to band in input extract (6% of starting material). A band was also detected in immunodepleted extract (6% of total unbound fraction), showing that the antibody does not completely deplete the antigen from cell lysate. In IF (1:200 ab dilution), a more intense staining was observed in WT cells whereas diffuse staining was observed in KO cells. Flow cytometry using 9532 (1:4380 ab dilution) showed a clear right-shift in WT cells relative to KO cells. KO cells also right shifted and some overlap between the two populations were observed.