Anti-PDE6 beta/PDE6B Antibody Picoband® (monoclonal, 8I2D7)

Western blot analysis of PDE6 beta/PDE6B using anti-PDE6 beta/PDE6B antibody (M02659-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: rat eye tissue lysates,
Lane 2: mouse eye 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 mouse anti-PDE6 beta/PDE6B antigen affinity purified monoclonal antibody (Catalog # M02659-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for PDE6 beta/PDE6B at approximately 98 kDa. The expected band size for PDE6 beta/PDE6B is at 98 kDa.

IHC analysis of PDE6 beta/PDE6B using anti-PDE6 beta/PDE6B antibody (M02659-1).
PDE6 beta/PDE6B was detected in a paraffin-embedded section of rat eye ball 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 μg/ml mouse anti-PDE6 beta/PDE6B Antibody (M02659-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

IF analysis of PDE6 beta/PDE6B using anti-PDE6 beta/PDE6B antibody (M02659-1).
PDE6 beta/PDE6B was detected in an immunocytochemical section of SH-SY5Y cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL mouse anti-PDE6 beta/PDE6B Antibody (M02659-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Flow Cytometry analysis of U20S cells using anti-PDE6 beta/PDE6B antibody (M02659-1).
Overlay histogram showing U20S cells stained with M02659-1 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with mouse anti-PDE6 beta/PDE6B Antibody (M02659-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.