Anti-Cyclophilin B PPIB Antibody Picoband® (monoclonal, 11C11)

Figure 1. Western blot analysis of Cyclophilin B using anti-Cyclophilin B antibody (M03229-1).
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 placenta tissue lysates,
Lane 2: U-87MG whole cell lysates,
Lane 3: HepG2 whole cell lysates,
Lane 4: Caco-2 whole cell lysates,
Lane 5: SW620 whole cell lysates,
Lane 6: PANC-1 whole cell lysates,
Lane 7: THP-1 whole cell lysates,
Lane 8: HEK293 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 mouse anti-Cyclophilin B antigen affinity purified monoclonal antibody (Catalog # M03229-1) 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 1:5000 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 Cyclophilin B at approximately 21KD. The expected band size for Cyclophilin B is at 21KD.

Figure 2. Western blot analysis of Cyclophilin B using anti-Cyclophilin B antibody (M03229-1).
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: rat spleen tissue lysates,
Lane 2: rat lung tissue lysates,
Lane 3: mouse spleen tissue lysates,
Lane 4: mouse lung tissue 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 mouse anti-Cyclophilin B antigen affinity purified monoclonal antibody (Catalog # M03229-1) 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 1:5000 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 Cyclophilin B at approximately 21KD. The expected band size for Cyclophilin B is at 21KD.

Figure 3. IHC analysis of Cyclophilin B using anti-Cyclophilin B antibody (M03229-1).
Cyclophilin B was detected in paraffin-embedded section of human oesophagus squama cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Cyclophilin B Antibody (M03229-1) 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.

Figure 4. IHC analysis of Cyclophilin B using anti-Cyclophilin B antibody (M03229-1).
Cyclophilin B was detected in paraffin-embedded section of human ovarian cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Cyclophilin B Antibody (M03229-1) 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.

Figure 5. IHC analysis of Cyclophilin B using anti-Cyclophilin B antibody (M03229-1).
Cyclophilin B was detected in paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Cyclophilin B Antibody (M03229-1) 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.

Figure 6. IHC analysis of Cyclophilin B using anti-Cyclophilin B antibody (M03229-1).
Cyclophilin B was detected in paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Cyclophilin B Antibody (M03229-1) 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.

Figure 7. IHC analysis of Cyclophilin B using anti-Cyclophilin B antibody (M03229-1).
Cyclophilin B was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Cyclophilin B Antibody (M03229-1) 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.

Figure 8. IHC analysis of Cyclophilin B using anti-Cyclophilin B antibody (M03229-1).
Cyclophilin B was detected in immunocytochemical section of U20S cell. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The cells were blocked with 10% goat serum. And then incubated with 2μg/ml mouse anti-Cyclophilin B Antibody (M03229-1) overnight at 4°C. Biotin conjugated goat anti-mouse IgG (BA1001) was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Cy3 Conjugated Avidin (BA1037). Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 9. Flow Cytometry analysis of U20S cells using anti-Cyclophilin B antibody (M03229-1).
Overlay histogram showing U20S cells stained with M03229-1 (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 mouse anti-Cyclophilin B Antibody (M03229-1,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse 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.