Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ HeLa cells and 4 µg of ab40888 [EP959Y]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.

Additional screenshots of mapped reads can be downloaded here.

Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol*. Cells were fixed with formaldehyde for 10min.
The  ChIP was performed with 25 µg of chromatin, 5 µg of  ab40888 (red), and 20 µl of Protein A/G sepharose beads. 5 μg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).
Primers and probes are located in the first kb of the transcribed region.

All lanes : Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888) at 1/20000 dilution (Purified)

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 500ng/ml Trichostatin A for 4 hours whole cell lysates

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 17 kDa
Observed band size: 17 kDa

All lanes : Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888) at 1/50000 dilution (Purified)

Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) treated with 500ng/ml Trichostatin A for 4 hours whole cell lysates
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 17 kDa
Observed band size: 17 kDa

All lanes : Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888) at 1/50000 dilution (Purified)

Lane 1 : C6 (Rat glial tumor glial cell) treated with 500ng/ml Trichostatin A for 4 hours whole cell lysates
Lane 2 : C6 (Rat glial tumor glial cell) whole cell lysates

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 17 kDa
Observed band size: 17 kDa

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human endometrium cancer tissue sections labeling Histone H3 with purified ab40888 at 1/4000 dilution (0.23 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) treated with 500ng/ml Trichostatin A for 4 hours cells labeling Histone H3 with purified ab40888 at 1/1000 dilution (0.9 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 (2.5 µg/ml) dilution. Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

All lanes : Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888) at 1/50000 dilution

Lane 1 : C6 cell lysate, untreated.
Lane 2 : C6 cell lysate, treated with TSA.

Lysates/proteins at 10 µg per lane.

Predicted band size: 17 kDa
Observed band size: 17 kDa
Additional bands at: 50 kDa (possible non-specific binding), 60 kDa (possible non-specific binding)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat stomach tissue sections labeling Histone H3 with purified ab40888 at 1/4000 dilution (0.23 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling Histone H3 with purified ab40888 at 1/4000 dilution (0.23 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

ab40888 diluted 1:100, staining acetylated histone H3 on human breast carcinoma sections.

ab40888 (1/500) staining Histone H3 (acetyl K18) in asynchronous HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X-100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.

See Abreview