DDC Antibody / DOPA decarboxylase (FY12305)

Flow Cytometry analysis of HepG2 cells using anti-DDC antibody. Overlay histogram showing HepG2 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-DDC 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.

Western blot analysis of DDC using anti-DDC antibody. Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human SHSY-5Y whole cell lysates, Lane 2: human Caco-2 whole cell lysates, Lane 3: human HepG2 whole cell lysates, Lane 4: rat NRK whole cell lysates, Lane 5: rat PC-12 whole cell lysates, Lane 6: mouse kidney tissue 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-DDC 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 an ECL Plus Western Blotting Substrate. The expected molecular weight of DDC is ~54 kDa.

Immunoprecipitation of DDC in SH-SY5Y whole cell lysate. Western blot analysis of DDC using anti-DDC antibody; Lane 1: SH-SY5Y whole cell lysates (30ug); Lane 2: Rabbit control IgG instead of anti-DDC antibody in SH-SY5Y whole cell lysate; Lane 3: anti-DDC antibody (2ug) + SH-SY5Y whole cell lysate (500ug). After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-DDC 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. The expected molecular weight of DDC is ~54 kDa.

Immunohistochemical staining of DDC using anti-DDC antibody. DDC was detected in a paraffin-embedded section of human kidney tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-DDC antibody overnight at 4oC. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37oC. The tissue section was developed using an HRP secondary and DAB substrate.