Galactosylceramidase (or galactocerebrosidase), EC 3.2.1.46, is an enzyme that removes galactose from ceramide derivatives (galactosylceramides) by catalysing the hydrolysis of galactose ester bonds of galactosylceramide, galactosylsphingosine, lactosylceramide, and monogalactosyldiglyceride. It is a lysosomal protein, encoded in humans by the GALC gene. Mutations in this gene have been associated with Krabbe disease, also known as galactosylceramide lipidosis.
Background References
1. Senkevich K et al. GALC variants affect galactosylceramidase enzymatic activity and risk of Parkinson\'s disease. Brain. 2023 May
2. Tian G et al. rAAV2-Mediated Restoration of GALC in Neural Stem Cells from Krabbe Patient-Derived iPSCs. Pharmaceuticals (Basel). 2023 Apr
Western blot analysis of GALC on different lysates with Rabbit anti-GALC antibody (HA722115) at 1/2,000 dilution.
Lane 1: 293T cell lysate (20 µg/Lane) Lane 2: A375 cell lysate (20 µg/Lane) Lane 3: A549 cell lysate (20 µg/Lane) Lane 4: SH-SY5Y cell lysate (20 µg/Lane)
Predicted band size: 77 kDa Observed band size: 48 kDa
Exposure time: 3 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722115) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-GALC antibody (HA722115) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722115) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue with Rabbit anti-GALC antibody (HA722115) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722115) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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