Anti-Histone H3 (acetyl K9) antibody - ChIP Grade

• Supplier: Abcam
• Catalog: ab10812
• Prices: $403.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: IHC-F IHC-P ChIP IP WB ChIPseq ICC/IF ICC/IF FC
• Species Reactivities: Mouse, Rat, Bovine, Human, Yeast, Arabidopsis thaliana, C. elegans, Drosophila, Zebrafish, Marmoset, Rice, Broad species
• Antigen: Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, acetylated at K9
• Blocking Peptide: Human Histone H3 (acetyl K9) peptide
• Description: Rabbit Polyclonal
• Gene: HIST1H3A
• Conjugate: Unconjugated

Product images

ab10812 staining histone H3 (acetyl K9) in A549 cells treated with scriptaid (ab120883), by ICC/IF. Increase in histone H3 (acetyl K9) expression correlates with increased concentration of scriptaid, as described in literature.The cells were incubated at 37°C for 24 hour in media containing different concentrations of ab120883 (scriptaid) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab10812 (0.1 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A anti-rabbit DyLight 488 secondary antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

ab10812 staining Histone H3 (acetyl K9) in Mouse thyroid tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.1% Tween 20 and blocked with 10% serum for 30 minutes at 24°C. Samples were incubated with primary antibody (1/200 in 10% serum in PBS) for 1 hour at 24°C. An Alexa Fluor® 555-conjugated Goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.See Abreview

All lanes : Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab10812) at 1 µg/mlLane 1 : Calf Thymus Histone Preparation Nuclear LysateLane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (unmodified ) peptide (ab2903) at 0.5 µg/mlLane 3 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (acetyl K9) peptide (ab16635) at 0.5 µg/mlLane 4 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H3 peptide - acetyl K14 at 0.5 µg/mlLane 5 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (acetyl K18) peptide (ab24003) at 0.5 µg/mlLane 6 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (acetyl K27) peptide (ab24404) at 0.5 µg/mlLane 7 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (acetyl K23) peptide (ab48359) at 0.5 µg/mlLysates/proteins at 0.5 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilutiondeveloped using the ECL techniqueP

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  ab10812 (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.     

ICC/IF image of ab10812 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 (ab10812, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 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. This antibody also gave a positive result in 100% Methanol fixed (5 min) Hek293, HepG2, and MCF-7 cells at 5µg/ml.

IHC image of Histone H3 (acetyl K9) 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 ab10812, 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.

SK-N-SH cells fixed with 4% paraformaldehyde, permeabilized in 0.5% Triton X-100 and incubated for 1 hour with ab10812 (1/1000). ab10812 clearly stained the nucleus (red). The cells were counterstained with DAPI (blue). 100x magnification.

ab10812 staining mouse embryonic stem cells by flow cytometry (gated on all living cells). The cells were incuabted with the antibody at 0.25ug/1.5 x 105 cells in a permeabilization buffer. A PE conjugated goat anti-rabbit antibody was used as the secondary.See Abreview

ab10812 staining Histone H3 (acetyl K9) in the root tips of Arabidopsis thaliana by Immunocytochemistry/ Immunofluorescence. Cells were fixed in 2% paraformaldehyde for 30 minutes at room temperature. Blocking and permeabilization was carried with 4% BSA solution containing 0,5% Triton X-100 in PBS at room temperature for 45 minutes. Slides were washed in PBS and incubated with primary antibody at a 1/200 dilution in 1% PBS for 1 hour at 37°C. Slides were washed in PBS and incubated with the secondary antibody ab6639 (Goat anti-rabbit Cy3 ® (H&L)) at a 1/500 dilution in 1% BSA in PBS for 1 hour at 37°C. Slides were counterstained with DAPI (2µg/mL) for 10 minutes at room temperature.Left image: DAPI stained interphase nucleus with prominent chromocentersMiddle image: Distribution of Histone H3 (acetyl K9) in the nucleus (arrows indicate absence of signal from chromocenters)Right image: Merged imageSee Abreview

All lanes : Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab10812) at 1/1000 dilutionLane 1 : Tissue lysate prepared from rat dorsal spinal cord at 5 µgLane 2 : Tissue lysate prepared from rat dorsal spinal cord at 5 µgLane 3 : Tissue lysate prepared from rat dorsal spinal cord at 8 µgLane 4 : Tissue lysate prepared from rat dorsal spinal cord at 8 µgLane 5 : Tissue lysate prepared from rat dorsal spinal cord at 8 µg with Human Histone H3 (acetyl K9) peptide (ab16635)SecondaryHRP conjugated rabbit polyclonaldeveloped using the ECL technique

Product References

Glycogen synthase kinase (GSK) 3beta phosphorylates and protects nuclear myosin - Glycogen synthase kinase (GSK) 3beta phosphorylates and protects nuclear myosin

Sarshad AA, Corcoran M, Al-Muzzaini B, Borgonovo-Brandter L, Von Euler A, Lamont D, Visa N, Percipalle P. PLoS Genet. 2014 Jun 5;10(6):e1004390.

Differential acetylation of histone H3 at the regulatory region of OsDREB1b - Differential acetylation of histone H3 at the regulatory region of OsDREB1b

Roy D, Paul A, Roy A, Ghosh R, Ganguly P, Chaudhuri S. PLoS One. 2014 Jun 18;9(6):e100343.

T cell activation regulates CD6 alternative splicing by transcription dynamics - T cell activation regulates CD6 alternative splicing by transcription dynamics

da Gloria VG, Martins de Araujo M, Mafalda Santos A, Leal R, de Almeida SF, Carmo AM, Moreira A. J Immunol. 2014 Jul 1;193(1):391-9.

Inhibition of DNA methylation alters chromatin organization, nuclear positioning - Inhibition of DNA methylation alters chromatin organization, nuclear positioning

Bockor VV, Barisic D, Horvat T, Maglica Z, Vojta A, Zoldos V. PLoS One. 2014 Aug 5;9(8):e103954.

A robust chromatin immunoprecipitation protocol for studying transcription - A robust chromatin immunoprecipitation protocol for studying transcription

Li W, Lin YC, Li Q, Shi R, Lin CY, Chen H, Chuang L, Qu GZ, Sederoff RR, Chiang VL. Nat Protoc. 2014 Sep;9(9):2180-93.

Pleckstrin homology domain leucine-rich repeat protein phosphatases set the - Pleckstrin homology domain leucine-rich repeat protein phosphatases set the

Reyes G, Niederst M, Cohen-Katsenelson K, Stender JD, Kunkel MT, Chen M, Brognard J, Sierecki E, Gao T, Nowak DG, Trotman LC, Glass CK, Newton AC. Proc Natl Acad Sci U S A. 2014 Sep 23;111(38):E3957-65. doi:

Alcohol consumption during gestation causes histone3 lysine9 hyperacetylation and - Alcohol consumption during gestation causes histone3 lysine9 hyperacetylation and

Pan B, Zhu J, Lv T, Sun H, Huang X, Tian J. Alcohol Clin Exp Res. 2014 Sep;38(9):2396-402.

Dose- and time-dependent epigenetic changes in the livers of Fisher 344 rats - Dose- and time-dependent epigenetic changes in the livers of Fisher 344 rats

Conti Ad, Kobets T, Escudero-Lourdes C, Montgomery B, Tryndyak V, Beland FA, Doerge DR, Pogribny IP. Toxicol Sci. 2014 Jun;139(2):371-80.

Active, phosphorylated fingolimod inhibits histone deacetylases and facilitates - Active, phosphorylated fingolimod inhibits histone deacetylases and facilitates

Hait NC, Wise LE, Allegood JC, O'Brien M, Avni D, Reeves TM, Knapp PE, Lu J, Luo C, Miles MF, Milstien S, Lichtman AH, Spiegel S. Nat Neurosci. 2014 Jul;17(7):971-80.