Anti-Histone H3 (acetyl K18) 抗体 - ChIP Grade

• WB

Ap

Western blot - Anti-Histone H3 (acetyl K18) antibody - ChIP Grade (AB1191)

All lanes:

Western blot - Anti-Histone H3 (acetyl K18) antibody - ChIP Grade (ab1191) at 1 µg/mL

Lane 1:

HeLa Histone Preparation Nuclear Lysate at 2.5 µg

Lane 2:

HeLa Histone Preparation Nuclear Lysate at 2.5 µg with Histone H3 peptide - unmodified R17

Lane 3:

HeLa Histone Preparation Nuclear Lysate at 2.5 µg with Human Histone H3 (acetyl K9) peptide (<a href='/products/proteins-peptides/human-histone-h3-acetyl-k9-peptide-ab16635'>ab16635</a>)

Lane 4:

HeLa Histone Preparation Nuclear Lysate at 2.5 µg with Histone H3 peptide - acetyl K14

Lane 5:

HeLa Histone Preparation Nuclear Lysate at 2.5 µg with Human Histone H3 (acetyl K18) peptide (ab24003)

Lane 6:

HeLa Histone Preparation Nuclear Lysate at 2.5 µg with Human Histone H3 (acetyl K23) peptide (<a href='/products/proteins-peptides/human-histone-h3-acetyl-k23-peptide-ab48359'>ab48359</a>)

Lane 7:

HeLa Histone Preparation Nuclear Lysate at 2.5 µg with Human Histone H3 (acetyl K27) peptide (<a href='/products/proteins-peptides/human-histone-h3-acetyl-k27-peptide-ab24404'>ab24404</a>)

Lane 8:

HeLa Histone Preparation Nuclear Lysate at 2.5 µg with Histone H3 peptide - acetyl K36

Lane 9:

HeLa Histone Preparation Nuclear Lysate at 2.5 µg with Human Histone H4 (acetyl K12) peptide (<a href='/products/proteins-peptides/human-histone-h4-acetyl-k12-peptide-ab15662'>ab15662</a>)

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 15 kDa

Observed band size: 17 kDa

true

Exposure time: 1min

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K18) antibody - ChIP Grade (AB1191)

IHC image of Histone H3 (acetyl K18) staining in Human normal skin formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab1191, 1μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K18) antibody - ChIP Grade (AB1191)

ab1191 staining Histone H3 (acetyl K18) in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab1191 at 0.1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 4% paraformaldehyde (10 min).Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ChIP

Unknown

ChIP - Anti-Histone H3 (acetyl K18) antibody - ChIP Grade (AB1191)

Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 μg of chromatin, 2 μg of ab1191 (blue), and 20 μl of protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.

• WB

Project

Western blot - Anti-Histone H3 (acetyl K18) antibody - ChIP Grade (AB1191)

All lanes:

Western blot - Anti-Histone H3 (acetyl K18) antibody - ChIP Grade (ab1191) at 1 µg/mL

Lane 1:

Histone prep at 0.5 µg/mL

Lane 2:

Histone prep at 0.5 µg/mL with Human Histone H3 (acetyl K18) peptide (ab24003)

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab6721'>ab6721</a>) at 1/5000 dilution

Predicted band size: 15 kDa

Observed band size: 17 kDa

false

• PepArr

Lab

Peptide Array - Anti-Histone H3 (acetyl K18) antibody - ChIP Grade (AB1191)

All batches of ab1191 are tested in Peptide Array against peptides to different Histone H3 and H4 modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - acetyl K18 peptide (ab24003), indicating that this antibody specifically recognises the Histone H3 - acetyl K18 modification.

ab17163 - Histone H3 - unmodified

ab16635 - Histone H3 - acetyl K9

ab15591 - Histone H3 - acetyl K14

ab24003 - Histone H3 - acetyl K18

ab48359 - Histone H3 - acetyl K23

ab24404 - Histone H3 - acetyl K27

ab41409 - Histone H3 - acetyl K36

ab15662 - Histone H4 - acetyl K12