Anti-Cyclophilin E/PPIE Antibody Picoband®

Western blot analysis of Cyclophilin E/PPIE using anti-Cyclophilin E/PPIE antibody (A08021-1).
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 50ug of sample under reducing conditions.
Lane 1: human HEK293 whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human PC-3 whole cell lysates,
Lane 5: human CACO-2 whole cell lysates,
Lane 6: mouse HEPA1-6 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-Cyclophilin E/PPIE antigen affinity purified polyclonal antibody (Catalog # A08021-1) 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-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 Cyclophilin E/PPIE at approximately 35KD. The expected band size for Cyclophilin E/PPIE is at 35KD.

IF analysis of Cyclophilin E/PPIE using anti-Cyclophilin E/PPIE antibody (A08021-1).
Cyclophilin E/PPIE was detected in immunocytochemical section of A431 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 5μg/mL rabbit anti-Cyclophilin E/PPIE Antibody (A08021-1) 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.

IF analysis of Cyclophilin E/PPIE using anti-Cyclophilin E/PPIE antibody (A08021-1).
Cyclophilin E/PPIE was detected in immunocytochemical section of PC-3 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 5μg/mL rabbit anti-Cyclophilin E/PPIE Antibody (A08021-1) 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.

Immunoprecipitating (IP) Cyclophilin E/PPIE in PC-3 whole cell lysate.
Western blot analysis of Cyclophilin E/PPIE using anti-Cyclophilin E/PPIE antibody (A08021-1);
Lane 1: PC-3 whole cell lysates (30ug);
Lane 2: Rabbit control IgG instead of anti-Cyclophilin E/PPIE antibody in PC-3 whole cell lysate;
Lane 3: anti-Cyclophilin E/PPIE antibody (2μg) + PC-3 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-Cyclophilin E/PPIE antigen affinity purified polyclonal antibody (A08021-1) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1196-200). A specific band was detected for Cyclophilin E/PPIE at approximately 35 kDa. The expected band size for Cyclophilin E/PPIE is at 35 kDa.

Flow Cytometry analysis of THP-1 cells using anti-Cyclophilin E/PPIE antibody (A08021-1).
Overlay histogram showing THP-1 cells stained with A08021-1 (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-Cyclophilin E/PPIE Antibody (A08021-1, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.