Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade

• Supplier: Abcam
• Catalog: ab8895
• Prices: $403.00
• Sizes: 50 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: IP FC Electron microscopy ICC/IF ICC/IF ICC/IF ChIP WB ChIPseq IHC-P ChIP ChIP
• Species Reactivities: Mouse, Human, Pig, Yeast, Xenopus, Drosophila, Plasmodium, Xenopus, Candida, Bovine, Deer, Plant, Mammalian
• Antigen: Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, mono methylated at K4
• Blocking Peptide: Human Histone H3 (mono methyl K4) peptide
• Description: Rabbit Polyclonal
• Gene: HIST3H3
• Conjugate: Unconjugated

Product images

ab8895 staining Histone H3 (mono methyl K4) in HeLa cells. The 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 ab8895 at 1µg/ml and ab7291 (anti beta Tubulin) at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat anti-rabbit AlexaFluor®488 secondary (ab150081) at 2 μg/ml (shown in green) and a goat anti-mouse AlexaFluor®594 secondary (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.Negative controls: 1– Rabbit primary antibody 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.

IHC image of ab8895 staining Histone H3 (mono methyl K4) in 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 ab8895, 0.5µg/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.

All lanes : Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895) (Sample: Calf thymus histone lysate)Lane 1 : As aboveLane 2 : Human Histone H3 (mono methyl K4) peptide (ab1340) at 1 µg/mlLane 3 : Human Histone H3 (di methyl K4) peptide (ab7768) at 1 µg/mlLane 4 : Human Histone H3 (tri methyl K4) peptide (ab1342) at 1 µg/mlLane 5 : Human Histone H3 (mono methyl K9) peptide (ab1771) at 1 µg/mlLane 6 : Human Histone H3 (mono methyl K27) peptide (ab1780) at 1 µg/mlLane 7 : Human Histone H3 (unmodified ) peptide (ab2903) at 1 µg/mlSecondaryGoat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilutionPerformed under reducing conditions.

Performed under reducing conditions.

Chromatin was prepared from U2OS 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  ab8895 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified on the GAPDH and ALDOA (active) and MYO-D (inactive) promoters and over the y-Actin gene (active). Schematic diagram of the y-Actin gene is shown on the top of the figure. Black boxes represent exons and thin lines represent introns. PCR products are depicted as bars under the gene.

A: Enlarged region showing the distribution of the H3K4me1 labelling around and within the EC cavities of a murine rod photoreceptor.B: Schematic representation of the labelling for the H3K4me1 mark. The bar represents 260 nm in A and 1 µm in B.

ab8895 used in Dot Blot at a 1/1000 dilution, 18 hours at 4°C.The dot blot was done using 0.1ug of peptide and screened multiple histone tail modifications to ensure no cross reactivity. See Abreview

Indian muntjac fibroblast cells -  interphase (top left) and prophase (top right and below), stained with:Mono Methyl K4 antibody, ab8895, (green) DAPI: red, top left and right; blue, belowPhospho Ser 10 antibody: red (below) and blue (top right)The perinuclear and perinucleolar heterochromatin domains do not contain Mono Methyl K4.  The Mono Methyl K4, rather, is distributed as small nuclear foci primarily found between DAPI-intense regions of the nucleus.   

ab8895 staining mouse embryonic stem cells by flow cytometry. The ES cell colonies were trypsinized and the cells permeabilized before being stained with ab8895 (0.25ug/1.5 x 105 cells). A PE conjugated goat anti-rabbit antibody was used as the secondary.See Abreview

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Biancolella M, Fortini BK, Tring S, Plummer SJ, Mendoza-Fandino GA, Hartiala J, Hitchler MJ, Yan C, Schumacher FR, Conti DV, Edlund CK, Noushmehr H, Coetzee SG, Bresalier RS, Ahnen DJ, Barry EL, Berman BP, Rice JC, Coetzee GA, Casey G. Hum Mol Genet. 2014 Apr 15;23(8):2198-209.

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Maroschik B, Gurtler A, Kramer A, Rossler U, Gomolka M, Hornhardt S, Mortl S, Friedl AA. Radiat Oncol. 2014 Jan 9;9:15.

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Looney TJ, Zhang L, Chen CH, Lee JH, Chari S, Mao FF, Pelizzola M, Zhang L, Lister R, Baker SW, Fernandes CJ, Gaetz J, Foshay KM, Clift KL, Zhang Z, Li WQ, Vallender EJ, Wagner U, Qin JY, Michelini KJ, Bugarija B, Park D, Aryee E, Stricker T, Zhou J, White KP, Ren B, Schroth GP, Ecker JR, Xiang AP, Lahn BT. Genome Res. 2014 Feb;24(2):267-80.

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Zentner GE, Balow SA, Scacheri PC. G3 (Bethesda). 2014 Feb 19;4(2):243-54.

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Denissov S, Hofemeister H, Marks H, Kranz A, Ciotta G, Singh S, Anastassiadis K, Stunnenberg HG, Stewart AF. Development. 2014 Feb;141(3):526-37.

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