Anti-Histone H3 (acetyl K27) antibody - ChIP Grade
| Name | Anti-Histone H3 (acetyl K27) antibody - ChIP Grade |
|---|---|
| Supplier | Abcam |
| Catalog | ab4729 |
| Prices | $404.00 |
| Sizes | 100 µg |
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Applications | IHC-F ICC/IF ICC/IF WB IHC-P ChIPseq ChIP ChIP ChIP Array |
| Species Reactivities | Mouse, Rat, Chicken, Bovine, Human, Arabidopsis thaliana, Drosophila, Monkey, Plasmodium, Rice, Xenopus |
| Antigen | Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, acetylated at K27 |
| Description | Rabbit Polyclonal |
| Gene | HIST1H3A |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
IHC image of ab4729 staining Histone H3 (acetyl K27) in normal human colon formalin-fixed paraffin-embedded tissue sections*, performed on a Leica Bond. 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 ab4729, 5µg/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. No primary antibody was used in the negative control (shown on the inset).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.*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
ab4729 staining Histone H3 (acetyl K27) in HeLa cells. The cells were incubated with 10mM Sodium butyrate (ab120948) for 6 hours (Treated) or solvent-only for control purposes (Non-treated). Cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab4729 at 0.5µg/ml and ab7291 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with a anti-rabbit AlexaFluor®488 secondary antibody (ab150077) at 2 μg/ml (shown in green) and a goat anti-mouse AlexaFluor®594 (ab150120) at 2 μg/ml (shown in pseudo colour red). Nuclear DNA was labelled in blue with DAPI.Negative controls: 1– Rabbit primary and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.
All batches of ab4729 are tested in Peptide Array against peptides to different Histone H3 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 K27 peptide (ab24404), indicating that this antibody specifically recognises the Histone H3 - acetyl K27 modification.ab24404 - Histone H3 - acetyl K27ab15591 - Histone H3 - acetyl K14ab24003 - Histone H3 - acetyl K18ab17163 - Histone H3 unmodifiedab48359 - Histone H3 - acetyl K23ab41409 - Histone H3 - acetyl K36ab15662 - Histone H4 - acetyl K12ab16635 - Histone H3 acetyl K9
developed using the ECL techniquePerformed under reducing conditions.
Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729) at 1 µg/ml + HeLa Histone Preparation Nuclear Lysate - Butyrate Treated at 2.5 µgSecondaryGoat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
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 25µg of chromatin, 2µg of ab4729 (blue), and 20µl 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.
IHC image of Histone H3 (acetyl K27) staining in human breast carcinoma FFPE section, performed on a 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 ab4729, 1µg/ml, for 8 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.
ab4729 (1/500) staining Histone H3 (acetyl K27) in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilized with 0.5% Triton X-100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please see abreview.See Abreview
Primary antibody: ab4729 (H3 acetyl K27)Dilution: 1/100ab4729 strongly stained histones of mouse ES cells. However, fluroescence was greatly diminished following pre-blocking using a H3 acetyl K9 peptide. This suggests the antibody cross-reacts with the K9 and K27 residues.
Lanes 1 & 3 : Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729) at 0.2 µg/mlLanes 2 & 4 : Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729) at 0.1 µg/mlLane 1 : Calf thymus histone lysateLane 2 : Calf thymus histone lysateLane 3 : Calf thymus histone lysate with Human Histone H3 (acetyl K27) peptide (ab24404) at 2 µgLane 4 : Calf thymus histone lysate with Human Histone H3 (acetyl K27) peptide (ab24404) at 2 µgLysates/proteins at 1 µg per lane.SecondaryGoat anti-rabbit (HRP) at 1/2000 dilution
All lanes : Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729) at 0.2 µg/mlLane 1 : HeLa whole cell lysateLane 2 : HeLa nuclear lysateLane 3 : HeLa whole cell lysate with Human Histone H3 peptide (ab17163) at 1 µg/mlLane 4 : HeLa nuclear lysate with Human Histone H3 peptide (ab17163) at 1 µg/mlLane 5 : HeLa whole cell lysate with Human Histone H3 (acetyl K27) peptide (ab24404) at 1 µg/mlLane 6 : HeLa nuclear lysate with Human Histone H3 (acetyl K27) peptide (ab24404) at 1 µg/mlLane 7 : HeLa whole cell lysate with Human Histone H3 (acetyl K9) peptide (ab16635) at 1 µg/mlLane 8 : HeLa nuclear lysate with Human Histone H3 (acetyl K9) peptide (ab16635) at 1 µg/mlLane 9 : HeLa whole cell lysate with Human Histone H3 (acetyl K18) peptide (ab24003) at 1 µg/mlLane 10 : HeLa nuclear lysate with Human Histone H3 (acetyl K18) peptide (ab24003) at 1 µg/mlLane 11 : HeLa whole cell lysate with Human Histone H3 (asymmetric di methyl R26) peptide (ab2854) at 1 µg/mlLane 12 : HeLa nuclear lysa
ICC/IF image of ab4729 stained Hela cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab4729, 1µg/ml) overnight at +4°C. The secondary antibody (green) was a goat anti-rabbit DyLight® 488 (IgG - H&L, pre-adsorbed) (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
ab4729 staining Histone H3 (acetyl K27) in Cyanidioschyzon merolae cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 5% BSA for 30 minutes at 37°C. Samples were incubated with primary antibody (1/100 in PBS + 0.1% BSA) for 1 hour at 37°C. An Alexa Fluor®488-conjugated Goat anti-rabbit polyclonal (100)was used as the secondary antibody.See Abreview
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