PARP-1, Abcam, ab32071

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).

In WB (1:1000 ab dilution) ab32071 detected a band of appr. 120 kDa in WT cell lysate with no corresponding band observed in KO cell lysate. 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 faint 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 (1:50 ab dilution), a more intense staining was observed in WT cell whereas KO cells exhibited some background staining. Flow cytometry using ab32071 (1:950 ab dilution) showed a pronounced right-shift in WT cells relative to KO cells. The KO population remained partly above the secondary-antibody control but was separated from WT cells, indicating substantial target-dependent staining with moderate background signal.