Anti-Epac1/RAPGEF3 Antibody Picoband®

Figure 1. Western blot analysis of Epac1/RAPGEF3 using anti-Epac1/RAPGEF3 antibody (A02483-3).
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: human Hela whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: monkey COS7 whole cell lysates,
Lane 4: human T-47D whole cell lysates,
Lane 5: mouse RAW264.7 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-Epac1/RAPGEF3 antigen affinity purified polyclonal antibody (Catalog # A02483-3) 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Epac1/RAPGEF3 at approximately 104 kDa. The expected band size for Epac1/RAPGEF3 is at 104 kDa.

Figure 2. IHC analysis of Epac1/RAPGEF3 using anti-Epac1/RAPGEF3 antibody (A02483-3).
Epac1/RAPGEF3 was detected in a paraffin-embedded section of human lung cancer 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 rabbit anti-Epac1/RAPGEF3 Antibody (A02483-3) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.

Figure 3. IHC analysis of Epac1/RAPGEF3 using anti-Epac1/RAPGEF3 antibody (A02483-3).
Epac1/RAPGEF3 was detected in a paraffin-embedded section of human pancreatic cancer 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 rabbit anti-Epac1/RAPGEF3 Antibody (A02483-3) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.

Figure 4. IHC analysis of Epac1/RAPGEF3 using anti-Epac1/RAPGEF3 antibody (A02483-3).
Epac1/RAPGEF3 was detected in a paraffin-embedded section of human rupture of spleen 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 rabbit anti-Epac1/RAPGEF3 Antibody (A02483-3) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.

Figure 5. IF analysis of Epac1/RAPGEF3 using anti-Epac1/RAPGEF3 antibody (A02483-3).
Epac1/RAPGEF3 was detected in an immunocytochemical section of T-47D 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 rabbit anti-Epac1/RAPGEF3 Antibody (A02483-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.

Figure 6. Flow Cytometry analysis of MCF-7 cells using anti-Epac1/RAPGEF3 antibody (A02483-3).
Overlay histogram showing MCF-7 cells stained with A02483-3 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-Epac1/RAPGEF3 Antibody (A02483-3, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit 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.