Anti Annexin A1/ANXA1

Anti-Annexin A1 antibody, PA1006, IHC(P)IHC(P): Human Placenta Tissue

Anti-Annexin A1 antibody, PA1006, IHC(P)IHC(P): Human Tonsil Tissue

Figure 4. IF analysis of Annexin A1/ANXA1 using anti-Annexin A1/ANXA1 antibody (PA1006).
Annexin A1/ANXA1 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-Annexin A1/ANXA1 Antibody (PA1006) 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 5. Flow Cytometry analysis of A431 cells using anti-Annexin A1/ANXA1 antibody (PA1006). Overlay histogram showing A431 cells stained with PA1006 (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 A1/ANXA1 Antibody (PA1006, 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 1. Western blot analysis of Annexin A1/ANXA1 using anti-Annexin A1/ANXA1 antibody (PA1006).
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 30ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human K562 whole cell lysates,
Lane 3: monkey COS-7 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-Annexin A1/ANXA1 antigen affinity purified polyclonal antibody (Catalog # PA1006) 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 Annexin A1/ANXA1 at approximately 39KD. The expected band size for Annexin A1/ANXA1 is at 39KD.