SCD1 Antibody / Stearoyl-CoA desaturase (FY12521)

Immunofluorescent staining of SCD using anti-SCD1 antibody (green). SCD1 was detected in an immunocytochemical section of cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/ml rabbit anti-SCD1 antibody overnight at 4oC. DyLight 488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37oC. The section was counterstained with DAPI nuclear stain (blue). Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of SCD1 using anti-SCD1 antibody. Lane 1: human whole cell lysates, Lane 2: human MCF-7 whole cell lysates, Lane 3: human HepG2 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-SCD antibody at 0.5 ug/ml overnight at 4oC, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal was developed using enhanced chemiluminescent. SCD1 (~41 kDa predicted) was detected as a 36-38 kDa doublet, consistent with the known anomalous migration and phosphorylation-dependent mobility of this endoplasmic reticulum membrane enzyme.

Flow Cytometry analysis of Caco-2 cells using anti-SCD1 antibody. Overlay histogram showing Caco-2 cells stained with (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-SCD1 antibody (1 ug/million cells) for 30 min at 20oC. DyLight 488 conjugated goat anti-rabbit IgG (5-10 ug/million cells) was used as secondary antibody for 30 minutes at 20oC. Isotype control antibody (Green line) was rabbit IgG (1 ug/million cells) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Immunoprecipitating SCD1 in whole cell lysate. Western blot analysis of SCD1 using anti-SCD antibody. Lane 1: whole cell lysates (30ug) Lane 2: Rabbit control IgG instead of anti-SCD antibody in whole cell lysate. Lane 3: anti-SCD1 antibody (2ug) + whole cell lysate (500ug) After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-SCD1 antibody at a dilution of 0.5 ug/ml and probed with a goat anti-rabbit IgG-HRP secondary antibody. The signal is developed using ECL Plus Western Blotting Substrate. SCD1 (~41 kDa predicted) was detected as a 36–38 kDa doublet, consistent with the known anomalous migration and phosphorylation-dependent mobility of this endoplasmic reticulum membrane enzyme.