Pro-cathepsin H, Abcam, ab88960

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). Furthermore, ab88960 was tested using HAP1, BxPC-3, Su-86.86, THP-1, AsPC-1 and FaDu cells in the application WB (WT cell lysates/100 µg protein).

In WB (1:1000 Ab dilution) ab88960 detected a band of appr. 25 kDa in HEK293T WT cell lysate with no corresponding band in HEK293T KO cell lysate. Stronger bands of appr. 50 and 70 kDa was observed for both genotypes. To compare protein levels in different cell types, WB (1:1000 Ab dilution) was done on HAP1, BxPC-3, Su-86.86, THP-1, AsPC-1 and FaDu WT cell lysates. HAP1 showed no protein level while all other cell types showed an intense band at appr. 25 kDa. 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.