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 (HA722004) at 1/5,000 dilution.
Lane 1: 293T cell lysate Lane 2: A375 cell lysate Lane 3: A549 cell lysate Lane 4: HepG2 cell lysate Lane 5: NIH/3T3 cell lysate Lane 6: PC-12 cell lysate Lane 7: Mouse brain tissue lysate Lane 8: Mouse testis tissue lysate Lane 9: Rat brain tissue lysate Lane 10: Rat testis tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 77 kDa Observed band size: 60 kDa
Exposure time: 43 seconds;
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 (HA722004) at 1/5,000 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.
Immunocytochemistry analysis of A549 cells labeling GALC with Rabbit anti-GALC antibody (HA722004) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-GALC antibody (HA722004) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"