Pro-cathepsin H, Thermo Fisher Scientific, MA5-17253

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Pro-cathepsin H 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 six commercial research antibodies against human Pro-cathepsin H encoded by CTSH.

The collection of antibodies was tested using HEK293T WT and CTSH KO cells in the applications WB (WT and KO cell lysates/100 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) MA5-17253 detected a band of appr. 25 kDa in WT cell lysate, with no corresponding band in KO cell lysate. Multiple intense bands of higher molecular weight were observed for both genotypes. A band was also detected following IP (2 µg Ab used). This band was 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) comparable staining intensities were observed in both KO and WT cells. Antibody performance was evaluated using the [F(WT)]/[F(KO)] ratio. A ratio of appr. 1 was observed, indicating no difference in fluorescence intensity between WT and KO cells.

We highly encourage other scientists to share their observations.