Anti-Cyclin E2/CCNE2 Antibody Picoband®

Western blot analysis of Cyclin E2/CCNE2 using anti-Cyclin E2/CCNE2 antibody (A04536-3).
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 Jurkat whole cell lysates,
Lane 2: human THP-1 whole cell lysates,
Lane 3: human MCF-7 whole cell lysates,
Lane 4: human T47D whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat C6 whole cell lysates,
Lane 7: mouse brain tissue lysates,
Lane 8: mouse Neuro-2a 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-Cyclin E2/CCNE2 antigen affinity purified polyclonal antibody (A04536-3) 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 Cyclin E2/CCNE2 at approximately 47 kDa. The expected band size for Cyclin E2/CCNE2 is at 47 kDa.

IHC analysis of Cyclin E2/CCNE2 using anti-Cyclin E2/CCNE2 antibody (A04536-3).
Cyclin E2/CCNE2 was detected in a paraffin-embedded section of mouse testis 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 2 μg/ml rabbit anti-Cyclin E2/CCNE2 Antibody (A04536-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IF analysis of Cyclin E2/CCNE2 using anti-Cyclin E2/CCNE2 antibody (A04536-3) and anti-Alpha Tubulin antibody (M03989-3).
Cyclin E2/CCNE2 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-Cyclin E2/CCNE2 Antibody (A04536-3) and mouse anti-Alpha Tubulin antibody (M03989-3) overnight at 4°C. Fluoro488 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.