WB
Western blot analysis of Histone H3 on different lysates with Mouse anti-Histone H3 antibody at 1/10,000 dilution.
Lane 1: HeLa cell lysate,
Lane 2: A549 cell lysate,
Lane 3: HT-29 cell lysate,
Lane 4: HEK-293 cell lysate,
Lane 5: C2C12 cell lysate,
Lane 6: L-929 cell lysate,
Lane 7: C6 cell lysate,
Lysates/proteins at 10 µg/Lane.
Exposure time: 30 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 1/10,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human testis tissue with Mouse anti-Histone H3 antibody at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 testis tissue with Mouse anti-Histone H3 antibody at 1/2,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 1/2,000 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 rat testis tissue with Mouse anti-Histone H3 antibody at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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.
mIHC
Fluorescence multiplex immunohistochemical analysis of mouse kidney (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-NPHS2 (Red), anti-Laminin beta 1 (Green), anti-Histone H3 (Blue), anti-SLC12A1 / NKCC2 (Cyan) and anti-CK18 (Magenta) on kidney. HRP Conjugated UltraPolymer Goat Polyclonal Antibody Mouse/Rabbit was used as a secondary antibody. The immunostaining was performed with the Five-Fluorescence immunohistochemical mouse/rabbit kit. The section was incubated in five rounds of staining: in the order of anti-NPHS2 (1/1,000 dilution), nti-Laminin beta 1 (1/1,000 dilution), anti-Histone H3(1/500 dilution),anti-SLC12A1 / NKCC2 (1/3,000 dilution) and anti-CK18 (1/3,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. Image acquisition was performed with Olympus VS200 Slide Scanner.
ChIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Histone H3 or Normal Mouse 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.