Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (AB177178)

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 Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178). 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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (AB177178)

Immunohistochemical analysis of paraffin-embedded human liver cancer tissue labeling Histone H3 (acetyl K27) with ab177178 at 1/1500 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Nuclear staining on human liver cancer tissue is observed.

The section was incubated with ab177178 for 30 minutes at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) was performed for 20 minutes.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (AB177178)

Immunohistochemical analysis of formalin fixed paraffin embedded human liver carcinoma labelling Histone H3 (acetyl K27) with ab177178 at a concentration of 0.1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab177178 Anti-Histone H3 (acetyl K27) antibody [EP16602] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (AB177178)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, labeling Histone H3 (acetyl K27) with ab177178 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing increased nuclear staining in HeLa cells treated with TSA (500 ng/ml, 4 hours).

The nuclear counter stain is DAPI (blue).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (AB177178)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) treated with 500 ng/ml Trichostatin A for 4 hours (Red)/untreated (Green) at 1/1500 dilution. The secondary antibody was Goat anti rabbit IgG (Alexa Fluor^®488) at 1/2000 dilution. Rabbit monoclonal IgG (ab172730) (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

• ChIP

Lab

ChIP - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (AB177178)

Chromatin was prepared from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25μg of chromatin, 2μg of ab177178 (blue), and 20μl of Protein A/G Sepharose beads. 2μg of rabbit normal IgG was added to the beads as a control sample (yellow). 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.

*http://www.abcam.com/resources?keywords=X%20ChIP%20protocol

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (AB177178)

Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling Histone H3 (acetyl K27) with ab177178 at 1/10000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Nuclear staining on mouse lung tissue is observed.

The section was incubated with ab177178 for 30 minutes at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) was performed for 20 minutes.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (AB177178)

Immunohistochemical analysis of paraffin-embedded rat pancreas tissue labeling Histone H3 (acetyl K27) with ab177178 at 1/10000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Nuclear staining on rat pancreas is observed.

The section was incubated with ab177178 for 30 minutes at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument. Counter stained with Hematoxylin.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) was performed for 20 minutes.

• WB

Unknown

Western blot - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (AB177178)

All lanes:

Western blot - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178) at 1/10000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) treated with 500 ng/ml Trichostatin A for 4 hours whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 15 kDa

Observed band size: 15 kDa

false

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (AB177178)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human epithelial cell line from cervix adenocarcinoma) cells and 5 µg of ab177178 [EP16602]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (AB177178)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human epithelial cell line from cervix adenocarcinoma) cells and 5 µg of ab177178 [EP16602]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (AB177178)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human epithelial cell line from cervix adenocarcinoma) cells and 5 µg of ab177178 [EP16602]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• WB

Unknown

Western blot - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (AB177178)

All lanes:

Western blot - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178) at 1/10000 dilution

Lane 1:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 15 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 500 ng/ml Trichostatin A for 4 hours whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 15 kDa

Observed band size: 15 kDa

false

• WB

Unknown

Western blot - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (AB177178)

All lanes:

Western blot - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178) at 1/10000 dilution

Lane 1:

C6 (Rat glial tumor glial cell) whole cell lysate at 15 µg

Lane 2:

C6 (Rat glial tumor glial cell) treated with 500 ng/ml Trichostatin A for 4 hours whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 15 kDa

Observed band size: 15 kDa

false

• PepArr

Unknown

Peptide Array - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (AB177178)

ab177178 was tested in Peptide array against 501 different modified and unmodified histone peptides; each peptide is printed on the array at six concentrations (each in triplicate).

Circle area represents affinity between the antibody and a peptide : all antigen-containing peptides are displayed as red circles, all other peptides as blue circles. The affinity is calculated as area under curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity.

The complete dataset, including full list of all peptides and information on the position of each peptide in the diagram, is available under the Product Protocol section.