Anti-ATRIP Antibody Picoband™

Figure 1. Western blot analysis of ATRIP using anti-ATRIP antibody (A03862-2).
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 MCF-7 whole cell lysates,
Lane 2: human PC-3 whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human HepG2 whole cell lysates,
Lane 5: monkey COS-7 whole cell lysates,
Lane 6: human Jurkat whole cell lysates,
Lane 7: human HEL whole cell lysates,
Lane 8: rat liver tissue lysates,
Lane 9: mouse lung tissue lysates,
Lane 10: mouse liver tissue lysates,
Lane 11: mouse RAW264.7 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-ATRIP antigen affinity purified polyclonal antibody (Catalog # A03862-2) at 0.25 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 ATRIP at approximately 86-90 kDa. The expected band size for ATRIP is at 86 kDa.

Figure 2. IF analysis of ATRIP using anti-ATRIP antibody (A03862-2).
ATRIP was detected in an immunocytochemical section of Hela 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 5 μg/mL rabbit anti-ATRIP Antibody (A03862-2) 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 3. Flow Cytometry analysis of HepG2 cells using anti-ATRIP antibody (A03862-2).
Overlay histogram showing HepG2 cells stained with A03862-2 (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-ATRIP Antibody (A03862-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.