WB
Western blot analysis of Histone H3 (acetyl K27) on different lysates with Rabbit anti-Histone H3 (acetyl K27) antibody at 1/2,000 dilution.
Lane 1: HeLa cell lysate,
Lane 2: HeLa treated with 500ng/mL TSA for 4 hours cell lysate,
Lane 3: NIH/3T3 cell lysate,
Lane 4: NIH/3T3 treated with 400nM TSA for 18 hours cell lysate,
Lane 5: C6 cell lysate,
Lane 6: C6 treated with 1μM TSA for 18 hours cell lysate,
Lysates/proteins at 20 µg/Lane.
Exposure time: 4 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
WB
Western blot analysis of Histone H3 (acetyl K27) on different lysates with Rabbit anti-Histone H3 (acetyl K27) antibody at 1/1,000 dilution.
Lane 1: Mouse brain tissue lysate,
Lane 2: Mouse hippocampus tissue lysate,
Lysates/proteins at 40 µg/Lane.
Exposure time: 4 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-Histone H3 (acetyl K27) antibody at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Histone H3 (acetyl K27) antibody at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ChIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with 500ng/mL TSA for 4 hours with Histone H3 (acetyl K27) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.