Galactosylceramidase, ABclonal, A23229

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, A23229 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 (1:1000 Ab dilution) A23229 detected the precursor form of GALC at appr. 74 kDa and detects the N-terminal cleaved form of GALC at appr. 50 kDa in OCUM-1 WT cell lysate. Bands not observed in OCUM-1 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, A23229 (1:1000 Ab dilution) detected a band of appr. 50 kDa in all cell types. Various amount of protein was observed between cell types. A faint band of appr. 74 kDa was also observed in AsPC-1, MCF7 and U-2 OS WT cell lysates. A band of ~74 kDa and ~48 kDa was detected following IP (2 µg Ab used). The 74 kDa and 48 kDa bands were more intense than the corresponding bands in input extract (5% of starting material). Bands 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. 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.