Anti-Histone H3 (acetyl K27) Rabbit Monoclonal Antibody

IHC analysis of Histone H3 (acetyl K27) using anti-Histone H3 (acetyl K27) antibody (M12477-15).
Histone H3 (acetyl K27) was detected in a paraffin-embedded section of human brain 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 1:200 rabbit anti-Histone H3 (acetyl K27) (M12477-15) overnight at 4°C. Polymer anti-rabbit IgG-HRP IHC detection kit was used as secondary antibody and incubated for 45 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.

Western blot analysis of Histone H3 using anti-Histone H3 antibody (M12477-15).
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 293T whole cell lysates,
Lane 3: human CACO-2 whole cell lysates,
Lane 4: human HepG2 whole cell lysates,
Lane 5: rat PC-12 whole cell lysates,
Lane 6: rat C6 whole cell lysates,
Lane 7: mouse RAW264.7 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-Histone H3 antigen affinity purified monoclonal antibody (Catalog # M12477-15) at 1:500 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 1:500 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 Histone H3 at approximately 15 kDa. The expected band size for Histone H3 is at 15 kDa.