Calpain-2 catalytic subunit, Thermo Fisher Scientific, PA5-27720

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Calpain-2 catalytic subunit 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 13 commercial research antibodies against human Calpain-2 catalytic subunit encoded by CAPN2.

The collection of antibodies was tested using MDA-MB-231 WT and CAPN2 KO cells in the applications WB (WT and KO cell lysates/20 µg protein), IP (WT cell lysate) and IF (WT and KO cells) according to YCharOS standard protocols (see reference).

In WB (1:1000 Ab dilution) PA5-27720 detected a band of appr. 74 kDa, with no corresponding band in KO cell lysate. An intense band of lower molecular weight was also detected in both genotypes. A band was also detected following IP (1 µg Ab used). This band was less 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), comparable staining intensities were observed in both KO and WT cells.

We highly encourage other scientists to share their observations.