Anti-Annexin V/ANXA5 Antibody Picoband®

Figure 1. IHC analysis of Annexin V using anti-Annexin V antibody (PB9044).
Annexin V was detected in paraffin-embedded section of Human Intestinal Cancer Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Annexin V Antibody (PB9044) 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 2. IHC analysis of Annexin V using anti-Annexin V antibody (PB9044).
Annexin V was detected in paraffin-embedded section of Human Mammary Cancer Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Annexin V Antibody (PB9044) 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 Annexin V using anti-Annexin V antibody (PB9044).
Annexin V was detected in paraffin-embedded section of Mouse Kidney Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Annexin V Antibody (PB9044) 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 Annexin V using anti-Annexin V antibody (PB9044).
Annexin V was detected in paraffin-embedded section of Rat Kidney Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Annexin V Antibody (PB9044) 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. Western blot analysis of Annexin V using anti-Annexin V antibody (PB9044).
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 Lung Tissue Lysate
Lane 2: Rat Brain Tissue Lysate
Lane 3: Rat Cardiac Muscle Tissue Lysate
Lane 4: Rat Kidney Tissue Lysate
Lane 5: HELA Whole Cell Lysate
Lane 6: SMMC Whole Cell Lysate
Lane 7: A549 Whole Cell Lysate
Lane 8: SGC Whole Cell Lysate
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-Annexin V antigen affinity purified polyclonal antibody (Catalog # PB9044) 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 1:10000 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 Annexin V at approximately 36KD. The expected band size for Annexin V is at 36KD.

Figure 6. IHC analysis of Annexin V using anti-Annexin V antibody (PB9044).
Annexin V was detected in frozen section of rat kidney tissues. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Annexin V Antibody (PB9044) 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 7. IHC analysis of Annexin V using anti-Annexin V antibody (PB9044).
Annexin V was detected in frozen section of human placenta tissues. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Annexin V Antibody (PB9044) 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 8. IF analysis of Annexin V using anti-Annexin V antibody (PB9044).
Annexin V was detected in immunocytochemical section of U20S cell. 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 2μg/mL rabbit anti-Annexin V Antibody (PB9044) 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 9. IF analysis of Annexin V using anti-Annexin V antibody (PB9044).
Annexin V was detected in immunocytochemical section of RH35 cell. 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 2μg/mL rabbit anti-Annexin V Antibody (PB9044) 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 10. Flow Cytometry analysis of PC-3 cells using anti-Annexin V antibody (PB9044).
Overlay histogram showing PC-3 cells stained with PB9044 (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-Annexin V Antibody (PB9044,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight488 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.

Figure 11. Flow Cytometry analysis of SiHa cells using anti-Annexin V antibody (PB9044).
Overlay histogram showing SiHa cells stained with PB9044 (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-Annexin V Antibody (PB9044,1μg/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.

Figure 12. IF analysis of Annexin V using anti-Annexin V antibody (PB9044)
Annexin V was detected in paraffin-embedded section of human placenta cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/mL rabbit anti-Annexin V Antibody (PB9044) overnight at 4°C. FITC Conjugated Goat Anti-Rabbit IgG (BA1105) 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.