Anti-POLDIP2 Antibody Picoband®

Western blot analysis of POLDIP2 using anti-POLDIP2 antibody (A09581-4).
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 Hela whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human Jurkat whole cell lysates,
Lane 4: human K562 whole cell lysates,
Lane 5: rat testis tissue lysates,
Lane 6: rat kidney tissue lysates,
Lane 7: mouse testis tissue lysates,
Lane 8: mouse kidney tissue 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-POLDIP2 antigen affinity purified polyclonal antibody (Catalog # A09581-4) 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 POLDIP2 at approximately 36 kDa. The expected band size for POLDIP2 is at 42 kDa.

Immunoprecipitating POLDIP2 in HepG2 whole cell lysate.
Western blot analysis of POLDIP2 using anti-POLDIP2 antibody (A09581-4);
Lane 1: HepG2 whole cell lysates (30ug);
Lane 2: Rabbit control IgG instead of anti-POLDIP2 antibody in HepG2 whole cell lysate;
Lane 3: anti-POLDIP2 antibody (2μg) + HepG2 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-POLDIP2 antigen affinity purified polyclonal antibody (A09581-4) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1196-200). A specific band was detected for POLDIP2 at approximately 38 kDa. The expected band size for POLDIP2 is at 42 kDa.

IF analysis of POLDIP2 using anti-POLDIP2 antibody (A09581-4).
POLDIP2 was detected in an immunocytochemical section of U2OS 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-POLDIP2 Antibody (A09581-4) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.

IF analysis of POLDIP2 using anti-POLDIP2 antibody (A09581-4).
POLDIP2 was detected in an immunocytochemical section of human SK-HEP-1 cells. Cells were permeabilized with Triton X-100 (AR0205) for 10 minutes. The cells were blocked with 10% goat serum. And then incubated with 10 μg/mL rabbit anti-POLDIP2 Antibody (A09581-4) overnight at 4°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127), Cy3 conjugated goat anti-mouse IgG (BA1031) 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.

Flow Cytometry analysis of U251 cells using anti-POLDIP2 antibody (A09581-4).
Overlay histogram showing U251 cells stained with A09581-4 (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-POLDIP2 Antibody (A09581-4, 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 (Red line) was also used as a control.