Anti-Histone H3 (acetyl K23) antibody [EPR17712] - ChIP Grade

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K23) antibody [EPR17712] - ChIP Grade (AB177275)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Histone H3 (acetyl K23) with ab177275 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. Acetylation level increased after treatment with Trichostatin A (500 ng/ml) for 4 hours. The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative controls are as follows : -
-ve control 1 : ab177275 at 1/2000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2 : Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• ChIP-seq

Lab

ChIP-sequencing - Anti-Histone H3 (acetyl K23) antibody [EPR17712] - ChIP Grade (AB177275)

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 ab177275 [EPR17712]. 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.

• ChIP

Supplier Data

ChIP - Anti-Histone H3 (acetyl K23) antibody [EPR17712] - ChIP Grade (AB177275)

Chromatin was prepared from HeLa (Human epithelial cells 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 ab177275 (blue), and 20μl of Anti Rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

• WB

Supplier Data

Western blot - Anti-Histone H3 (acetyl K23) antibody [EPR17712] - ChIP Grade (AB177275)

Blocking/Dilution buffer : 5% BSA/TBST.

All lanes:

Western blot - Anti-Histone H3 (acetyl K23) antibody [EPR17712] - ChIP Grade (ab177275) at 1/1000 dilution

Lane 1:

Untreated HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 10 µg

Lane 2:

HeLa (Human epithelial cells from cervix adenocarcinoma) treated with 500 ng/ml Trichostatin A for 4 hours whole cell lysate at 10 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 1/100000 dilution

Predicted band size: 15 kDa

Observed band size: 15 kDa

false

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-Histone H3 (acetyl K23) antibody [EPR17712] - ChIP Grade (AB177275)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Histone H3 (acetyl K23) antibody [EPR17712] - ChIP Grade (ab177275) at 1/1000 dilution

Lane 1:

Untreated NIH/3T3 (Mouse embryonic fibroblast cells) whole cell lysate at 10 µg

Lane 2:

NIH/3T3 (Mouse embyronic fibroblast cells) treated with 500 ng/ml Trichostatin A for 4 hours whole cell lysate at 10 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 1/100000 dilution

Predicted band size: 15 kDa

Observed band size: 15 kDa

false

Exposure time: 3min

• PepArr

Supplier Data

Peptide Array - Anti-Histone H3 (acetyl K23) antibody [EPR17712] - ChIP Grade (AB177275)

ab177275 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.