Anti-Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade

• Supplier: Abcam
• Catalog: ab32356
• Prices: $404.00
• Sizes: 100 µl
• Host: Rabbit
• Clonality: Monoclonal
• Isotype: IgG
• Clone: Y47
• Applications: ChIPseq IHC-F ChIP WB IHC-P ICC/IF ICC/IF IP FC
• Species Reactivities: Mouse, Rat, Human, Monkey, Rice, Sheep, Chicken, Bovine, Yeast, Xenopus, Arabidopsis thaliana, Zebrafish, Mammalian
• Antigen: A synthetic (Dimethyl K) peptide corresponding to residues surrounding Lys4 of Histone H3
• Description: Rabbit Monoclonal
• Gene: HIST3H3
• Conjugate: Unconjugated

Product images

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, 8µl of ab32356 (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.

Ab32356, at a 1/250 dilution, staining Histone H3 in HeLa cells by Immunofluorescence.

ab32356 (1/200) staining Histone H3 (di methyl K4) in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilized with 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus. For further experimental details please refer to Abreview.See Abreview

ICC/IF image of ab32356 stained HepG2 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 (ab32356 , 5µg/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.

Overlay histogram showing HeLa cells stained with ab32356 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32356, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was a goat anti-rabbit DyLight® 488 (IgG; H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

All lanes : Anti-Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade (ab32356) at 1/20000 dilutionLane 1 : HeLa cell lysateLane 2 : Non methylated recombinant Histone H3

Ab32356, at a 1/100 dilution, staining Histone H3 in paraffin embedded breast carcinoma tissue sections by Immunohistochemistry. Ab32356, at a 1/100 dilution, staining Histone H3 in paraffin embedded breast carcinoma tissue sections by Immunohistochemistry.

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