Anti-Histone H4 (acetyl K12) 抗体 [EPR28340-173]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 (acetyl K12) antibody [EPR28340-173] (AB320815)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Histone H4 (acetyl K12) with ab320815 at 1/500 (1.044 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/mL dilution (Green).

Confocal image showing increased nuclear staining in HeLa cells treated with Trichostatin A (500 ng/mL) for 4 hr (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

• Flow Cyt

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Flow Cytometry - Anti-Histone H4 (acetyl K12) antibody [EPR28340-173] (AB320815)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) treated with 500ng/ml Trichostatin A for another 4h (Green) /Untreated control (Magenta) cells labelling Histone H4 (acetyl K12) with ab320815 at 1/50 dilution (1ug) / magenta and green compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488) (ab199091) (Black) and Grey isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ChIP-seq

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ChIP-sequencing - Anti-Histone H4 (acetyl K12) antibody [EPR28340-173] (AB320815)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 4 µg of ab320815 [EPR28340-173]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 60 million reads. The Input control is also shown.

• ChIP-seq

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ChIP-sequencing - Anti-Histone H4 (acetyl K12) antibody [EPR28340-173] (AB320815)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 4 µg of ab320815 [EPR28340-173]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 60 million reads. The Input control is also shown.

• IP

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Immunoprecipitation - Anti-Histone H4 (acetyl K12) antibody [EPR28340-173] (AB320815)

Histone H4 (acetyl K12) was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) treated with 500ng/ml TSA for 4h whole cell lysate with ab320815 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab320815 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) treated with 500ng/ml TSA for 4h whole cell lysate 10ug
Lane 2 : ab320815 IP in HeLa (human cervical adenocarcinoma epithelial cell) treated with 500ng/ml TSA for 4h whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab320815 in HeLa treated with 500ng/ml TSA for 4h whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST

The IP experiment was performed by ab320815 (capture antibody), and band detected by another Anti-Histone H4 (acetyl K12) antibody [EPR17906] (ab177793).

All lanes:

Immunoprecipitation - Anti-Histone H4 (acetyl K12) antibody [EPR28340-173] (ab320815) at 1/30 dilution

All lanes:

HeLa (human cervical adenocarcinoma epithelial cell) treated with 500ng/ml TSA for 4h whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 67s

• ChIP-seq

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ChIP-sequencing - Anti-Histone H4 (acetyl K12) antibody [EPR28340-173] (AB320815)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 4 µg of ab320815 [EPR28340-173]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 60 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-Histone H4 (acetyl K12) antibody [EPR28340-173] (AB320815)

Chromatin was prepared from NIH/3T3 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 4 µg of ab320815 [EPR28340-173]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 60 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-Histone H4 (acetyl K12) antibody [EPR28340-173] (AB320815)

Chromatin was prepared from NIH/3T3 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 4 µg of ab320815 [EPR28340-173]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 60 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-Histone H4 (acetyl K12) antibody [EPR28340-173] (AB320815)

Chromatin was prepared from NIH/3T3 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 4 µg of ab320815 [EPR28340-173]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 60 million reads. The Input control is also shown.

• PepArr

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Peptide Array - Anti-Histone H4 (acetyl K12) antibody [EPR28340-173] (AB320815)

Peptide array analysis of ab320815 at 1 : 5000 (0.1 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Fluo 647nm conjugated at 1 : 50,000 dilution.

ab320815 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 the area under the 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 a full list of all peptides and information on the position of each peptide in the diagram, can be downloaded here.