Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade

• Supplier: Abcam
• Catalog: ab8896
• Prices: $401.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: WB ChIP ICC/IF ICC/IF ChIPseq
• Species Reactivities: Bovine, Human, C. elegans, Broad species, Mammalian
• Antigen: Synthetic peptide derived from within residues 1 - 100 Histone H3
• Description: Rabbit Polyclonal
• Gene: HIST3H3
• Conjugate: Unconjugated

Product images

All lanes : Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab8896) at 1 µg/mlLane 1 : Calf Thymus Histone Preparation Lane 2 : Calf Thymus Histone Preparation with Human Histone H3 (mono methyl K9) peptide (ab1771) at 0.5 µg/mlLane 3 : Calf Thymus Histone Preparation with Human Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µg/mlLane 4 : Calf Thymus Histone Preparation with Human Histone H3 (tri methyl K9) peptide (ab1773) at 0.5 µg/mlLane 5 : Calf Thymus Histone Preparation with Human Histone H3 (mono methyl K27) peptide (ab1780) at 0.5 µg/mlLane 6 : Calf Thymus Histone Preparation with Human Histone H3 (di methyl K27) peptide (ab1781) at 0.5 µg/mlLane 7 : Calf Thymus Histone Preparation with Human Histone H3 (tri methyl K27) peptide (ab1782) at 0.5 µg/mlLane 8 : Calf Thymus Histone Preparation with Human Histone H3 (mono methyl K4) peptide (ab1340) at 0.5 µg/mlLane 9 : Calf Thymus Histone Preparation with Human Histone H3 (unmodified ) peptide (ab7228) at 0.5 µg/mlLane 10

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  ab8896 (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 for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.

ab8896 staining Histone H3 (mono methyl K9) in human differentiated haematopoietic stem cells by Flow Cytometry. Cells were fixed with paraformaldehyde and permeabilized with permeabilization buffer. The sample was incubated with the primary antibody (1/600) for 12 hours at 4°C. An Alexa Fluor® 488-conjugated goat polyclonal anti-rabbit IgG (1/1000) was used as the secondary antibody.Gating Strategy: Isotype negative control (white).See Abreview

ICC/IF image of ab8896 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 ab8896 at 1µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was a goat anti-rabbit Alexa Fluor® 488 (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

All lanes : Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab8896) at 1/1000 dilutionLane 1 : Nuclear lysates prepared from Hela cell: SiRNA untreatedLane 2 : Nuclear lysates prepared from Hela cells treated with G9a SiRNALysates/proteins at 10 µg per lane.SecondaryAlkaline Phosphatase conjugated rabbit polyclonal rabbit IgG (Fc) at 1/5000 dilution

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