Anti-PCYT2 Antibody Picoband®

Western blot analysis of PCYT2 using anti-PCYT2 antibody (A07910-1).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human HepG2 whole cell lysates,
Lane 2: human RT4 whole cell lysates,
Lane 3: human Caco-2 whole cell lysates,
Lane 4: human K562 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: rat RH35 whole cell lysates,
Lane 7: mouse liver tissue lysates,
Lane 8: mouse HEPA1-6 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-PCYT2 antigen affinity purified polyclonal antibody (A07910-1) 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 ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for PCYT2 at approximately 44 kDa. The expected band size for PCYT2 is at 44 kDa.

IF analysis of PCYT2 using anti-PCYT2 antibody (A07910-1) and anti-Beta Tubulin antibody (M01857-3).
PCYT2 was detected in an immunocytochemical section of HepG2 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-Histone H4 Antibody (A07910-1) and mouse anti-PCYT2 antibody (M01857-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) and Cy3 Conjugated Goat Anti-Mouse IgG (BA1031) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Immunoprecipitating (IP) PCYT2 in K562 whole cell lysate.
Western blot analysis of PCYT2 using anti-PCYT2 antibody (A07910-1);
Lane 1: K562 whole cell lysates (30ug);
Lane 2: Rabbit control IgG instead of anti-PCYT2 antibody in K562 whole cell lysate;
Lane 3: anti-PCYT2 antibody (2μg) + K562 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-PCYT2 antigen affinity purified polyclonal antibody (A07910-1) 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 PCYT2 at approximately 42 kDa. The expected band size for PCYT2 is at 42 kDa.

Flow Cytometry analysis of K562 cells using anti-PCYT2 antibody (A07910-1).
Overlay histogram showing K562 cells stained with A07910-1 (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-PCYT2 Antibody (A07910-1, 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.