Galactosylceramidase, Abcam, ab240638

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).

In WB (1:500 Ab dilution) ab240638 detected multiple bands , none of which were specific to WT lysate. A band of ~74 kDa and ~48 kDa was detected following IP (2 µg Ab used). The 48 kDa band was 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 74 kDa had same intensity as band in input extract and immunodepleted extract. In IF (1:300 Ab dilution), staining of WT cells were more pronounced compared to KD cells which showed some background staining.

We highly encourage other scientists to share their observations.