Galactosylceramidase/GALC Antibody (PSH03-45)

Immunocytochemistry/ Immunofluorescence: Galactosylceramidase/GALC Antibody (PSH03-45) [NBP3-32361]

Immunocytochemistry/ Immunofluorescence: Galactosylceramidase/GALC Antibody (PSH03-45) [NBP3-32361] - Immunocytochemistry analysis of A549 cells labeling Galactosylceramidase/GALC with Rabbit anti-Galactosylceramidase/GALC antibody (NBP3-32361) 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-Galactosylceramidase/GALC antibody (NBP3-32361) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.

Detection of Galactosylceramidase/GALC by Immunoprecipitation.

OCUM-1 cell line lysates were prepared and immunoprecipitation was performed using 2.0 μg of Galactosylceramidase/GALC Antibody (PSH03-45) (Catalog # NBP3-32361) pre-coupled to Dynabeads Protein A. Immunoprecipitated Galactosylceramidase/GALC was detected in Western Blot with a rabbit Galactosylceramidase/GALC antibody used at 1/1000. The Ponceau stained transfer of the blot is shown. SM=4% starting material; UB=4% unbound fraction; IP=immunoprecipitate; HC=antibody heavy chain. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).

Western Blot: Galactosylceramidase/GALC Antibody (PSH03-45) [NBP3-32361]

Western blot analysis on different lysates with NBP3-32361 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. NBP3-32361 at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG [HRP] Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.