Anti-Histone H4 (di methyl K20, tri methyl K20) antibody [6F8-D9]

• Supplier: Abcam
• Catalog: ab78517
• Prices: $403.00
• Sizes: 100 µg
• Host: Mouse
• Clonality: Monoclonal
• Isotype: IgG1
• Clone: 6F8-D9
• Applications: IHC-P WB ELISA DB ICC/IF ICC/IF FC
• Species Reactivities: Mouse, Human, C. elegans, Xenopus, Arabidopsis thaliana, Drosophila, Mammalian
• Antigen: Synthetic peptide of Human Histone H4
• Description: Mouse Monoclonal
• Gene: His4:CG31611
• Conjugate: Unconjugated

Product images

ICC/IF image of ab78517 stained mouse oocyte. Oocytes were fixed in formaldehyde and then incubated in 3% serum with 0.25% Triton for 30 minutes at 37°C. Cells were incubated with the antibody (ab78517) at 1/200 for 2 hours at 37°C. The secondary antibody used was a polyclonal Goat-anti-mouse conjugated to Alexa Fluor® 488 (1/250).See Abreview

All lanes : Anti-Histone H4 (di methyl K20, tri methyl K20) antibody [6F8-D9] (ab78517) at 1 µg/mlLane 1 : Calf Thymus Histone Preparation Nuclear LysateLane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H4 peptide (ab14963) at 0.5 µg/mlLane 3 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K4) peptide (ab1340) at 0.5 µg/mlLane 4 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K4) peptide (ab7768) at 0.5 µg/mlLane 5 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K4) peptide (ab1342) at 0.5 µg/mlLane 6 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H4 (mono methyl K20) peptide (ab17043) at 0.5 µg/mlLane 7 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H4 peptide (17-24) - di methyl K20 at 0.5 µg/mlLane 8 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H4 (tri methyl K20) peptide (ab17567) at 0.5 µg/mlLan

ICC/IF image of ab78517 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 (ab78517, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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 MCF7 cells at 5µg/ml, and in 4% PFA fixed (10 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml.

IHC image of Histone H4 (tri methyl K20) staining in normal human skin formalin fixed paraffin embedded tissue section, performed on a Leica 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 ab78517, 1µg/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.

Overlay histogram showing HeLa cells stained with ab78517 (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. The cells were then incubated with the antibody (ab78517, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed.

Product References

Dynamics of 5-methylcytosine and 5-hydroxymethylcytosine during germ cell - Dynamics of 5-methylcytosine and 5-hydroxymethylcytosine during germ cell

Yamaguchi S, Hong K, Liu R, Inoue A, Shen L, Zhang K, Zhang Y. Cell Res. 2013 Mar;23(3):329-39.

Visualization of multivalent histone modification in a single cell reveals highly - Visualization of multivalent histone modification in a single cell reveals highly

Hattori N, Niwa T, Kimura K, Helin K, Ushijima T. Nucleic Acids Res. 2013 Aug;41(15):7231-9.

A non-canonical role for the C. elegans dosage compensation complex in growth and - A non-canonical role for the C. elegans dosage compensation complex in growth and

Webster CM, Wu L, Douglas D, Soukas AA. Development. 2013 Sep;140(17):3601-12.