Galactosylceramidase, GeneTex, GTX101197

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human galactosylceramidase 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 6 commercial research antibodies against human galactosylceramidase encoded by GALC gene.

The collection of antibodies was tested using OCUM-1 WT and GALC KD cells in the applications WB (WT and KD cell lysates/30 µg protein), IP (WT cell lysate) and IF (WT and KD cells) according to YCharOS standard protocols (see reference). Furthermore, GTX101197 was tested using AsPC-1, OCUM-1, SU.86.86, Daoy, HCT-15, MCF7, U-2 OS and PANC-1 WT cells in the application WB (WT and KD cell lysate/30 µg protein).

In WB (1:500 Ab dilution) GTX101197 detected the precursor form of GALC was detected at appr. 74 kDa and the C-terminal cleaved form at appr. 26 kDa in WT cell lysate not observed in KD cell lysate. A band of higher molecular weight was observed in both WT and KD cell lysate. In WB of AsPC-1, OCUM-1, SU.86.86, Daoy, HCT-15, MCF7, U-2 OS and PANC-1 WT cells, GTX101197 (1:500 Ab dilution) detected a band of appr. 135 kDa in all cell types, a band of appr. 74 kDa in OCUM-1 and MCF7, and a band of appr. 26 kDa in AxPC-1, OCUM-1, MCF7 and U-2 OS WT cell lysate. A band of ~74 kDa and ~26 kDa was detected following IP (2 µg Ab used). The 74 kDa band was markedly more intense than the corresponding band in input extract (5% of starting material). Band was also detected in immunodepleted extract (5% of total unbound fraction), showing that the antibody does not completely deplete the antigen from the cell lysate. The band of 26 kDa had same intensity as band in input extract and immunodepleted extract. 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.