ABCE1 Antibody

Flow Cytometry analysis of U251 cells using anti-ABCE1 antibody. Overlay histogram showing U251 cells (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-ABCE1 Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IF analysis of ABCE1 using anti-ABCE1 antibody. ABCE1 was detected in an immunocytochemical section of Hela cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL rabbit anti-ABCE1 Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG 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.

Immunoprecipitating ABCE1 in A431 whole cell lysate. Western blot analysis of ABCE1 using anti-ABCE1 antibody. Lane 1: A431 whole cell lysates (30 ug), Lane 2: Rabbit control IgG instead of anti-ABCE1 antibody in A431 whole cell lysate, Lane 3: anti-ABCE1 antibody (2 µg) + A431 whole cell lysate (500 µg). After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-ABCE1 antigen affinity purified polyclonal antibody at a dilution of 0.5 µg/mL and probed with a mouse anti-rabbit IgG-HRP secondary antibody (Heavy Chain). The signal is developed using ECL Plus Western Blotting Substrate. A specific band was detected for ABCE1 at approximately 67 kDa. The expected band size for ABCE1 is at 67 kDa.

Western blot analysis of ABCE1 using anti-ABCE1 antibody. 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 HepG2 whole cell lysates, Lane 2: human A431 whole cell lysates, Lane 3: human U2OS whole cell lysates, Lane 4: human Jurkat whole cell lysates, Lane 5: rat brain tissue lysates, Lane 6: mouse brian tissue lysates, Lane 7: mouse 4T1 whole cell lysates, Lane 8: mouse NIH/3T3 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-ABCE1 antigen affinity purified polyclonal antibody 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 with Tanon 5200 system. A specific band was detected for ABCE1 at approximately 67 kDa. The expected band size for ABCE1 is at 67 kDa.