Goat Anti-Rabbit IgG (H+L) Secondary Antibody, Fluoro647 Conjugated

Figure 1. Western blot analysis of APEX1 using anti-APEX1 antibody (PB9128).
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 K562 whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human Raji whole cell lysates,
Lane 4: human A431 whole cell lysates,
Lane 5: human A549 whole cell lysates,
Lane 6: human MCF-7 whole cell lysates,
Lane 7: human RT4 whole cell lysates,
Lane 8: human U87 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-APEX1 antigen affinity purified polyclonal antibody (Catalog # PB9128) 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-Fluoro647 Conjugated secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. A specific band was detected for APEX1 at approximately 36 kDa. The expected band size for APEX1 is at 36 kDa.

IF analysis of COX5B using anti-COX5B antibody (A06090-2).
COX5B was detected in a paraffin-embedded section of human intestinal 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 5 μg/mL rabbit anti-COX5B Antibody (A06090-2) overnight at 4°C. Fluoro647 Conjugated Goat Anti-Rabbit IgG (BA1150) was used as secondary antibody at 1:500 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.