Cathepsin b, R&D Systems, AF953

YCharOS previously conducted a large head-to-head study comparing commercial research antibodies against human Cathepsin B, encoded by CTSB. The antibodies were evaluated based on published scientific literature and experimental data according to the pAbmAbs rating criteria.

In the previous study, this antibody was evaluated by YCharOS using HAP1 WT and CTSB KO cells in Western blot (WB), following YCharOS standard protocols (see reference).

Building on these previous findings, the antibody was further evaluated by YCharOS in immunofluorescence (IF) using HAP1 WT and CTSB KO cells and in IP using HAP1 WT cell lysate. At an antibody dilution of 1:200, comparable staining intensity was observed between the WT and CTSB KO cells in IF, indicating no apparent target-specific staining under the tested conditions. A band was detected following IP (2 µg Ab used), this band was markedly more intense compared to band in input extract (4% of starting material) and no band was detected in immunodepleted extract (4% of total unbound fraction), showing that the antibody does completely deplete the antigen from cell lysate.

We highly encourage other scientists to share their experience.

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Cathepsin B 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 Cathepsin B encoded by CTSB.

The collection of antibodies was tested using HAP1 WT and CTSB KO cells in the application WB (WT and KO cell lysates/30 µg protein), according to YCharOS standard protocols (see reference).

In WB (1:1000 Ab dilution) AF953 detected no target protein, but a protein of higher molecular weight for both genotypes.

We highly encourage other scientists to share their observations.