Anti-Histone H3 (di+tri methyl K4) antibody [mAbcam 6000] - ChIP Grade
| Name | Anti-Histone H3 (di+tri methyl K4) antibody [mAbcam 6000] - ChIP Grade |
|---|---|
| Supplier | Abcam |
| Catalog | ab6000 |
| Prices | $404.00 |
| Sizes | 100 µg |
| Host | Mouse |
| Clonality | Monoclonal |
| Isotype | IgG2b |
| Clone | mAbcam 6000 |
| Applications | ICC/IF ICC/IF IHC-P WB ICC/IF ChIP FC |
| Species Reactivities | Mouse, Rabbit, Bovine, Human, Yeast, Drosophila, Rice, Candida, Rat, Chicken, Xenopus, Arabidopsis thaliana, C. elegans, S. pombe, Neurospora crassa |
| Antigen | Synthetic peptide derived from residues 1 - 100 of Human Histone H3, tri methylated at K4 |
| Description | Mouse Monoclonal |
| Gene | HIST3H3 |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
Immunofluorescent imaging of human cells (U2OS) with ab6000 reveals a diffuse background nuclear staining corresponding to global dimethylation of K4, with multiple foci of brighter staining. The complete lack of nucleolar or cytoplasmic staining background confirms the high specificity of this antibody in this application. IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour. Primary antibody used at 1/200 in 5% milk / 0.2% TWEEN for one hour, secondary antibody Alexa 488 for 30 minutes. All blocking and incubation steps carried out at 37 degrees.Due to the size constraints for images on our website, unfortunately we are unable to show a higher quality image.
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 25 µg of chromatin, 2 µg of ab6000 (blue), and 20 µl 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.
![All lanes : Anti-Histone H3 (di+tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000) at 1 µg/mlLane 1 : Calf Thymus Histone Preparation Nuclear LysateLane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (unmodified ) peptide (ab7228) at 0.5 µgLane 3 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K4) peptide (ab1340) at 0.5 µgLane 4 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K4) peptide (ab7768) at 0.5 µgLane 5 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K4) peptide (ab1342) at 0.5 µgLane 6 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K9) peptide (ab1771) at 0.5 µgLane 7 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µgLane 8 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K9) peptide (ab1773) at 0.5 µgLane](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/1708_ab6000-221321-WBAP17803311.jpg)
All lanes : Anti-Histone H3 (di+tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000) at 1 µg/mlLane 1 : Calf Thymus Histone Preparation Nuclear LysateLane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (unmodified ) peptide (ab7228) at 0.5 µgLane 3 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K4) peptide (ab1340) at 0.5 µgLane 4 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K4) peptide (ab7768) at 0.5 µgLane 5 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K4) peptide (ab1342) at 0.5 µgLane 6 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K9) peptide (ab1771) at 0.5 µgLane 7 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µgLane 8 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K9) peptide (ab1773) at 0.5 µgLane
![Overlay histogram showing HeLa cells stained with ab6000 (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 (ab6000, 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 IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/1709_Histone-H3-Primary-antibodies-ab6000-8.jpg)
Overlay histogram showing HeLa cells stained with ab6000 (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 (ab6000, 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 IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Left image shows human Hep cell monolayer fixed with 4% formaldehyde, permeabilized using 0.5% Triton X-100, blocked with 5% FCS in PBS (20 min) and incubated with ab6000 for 1 hr (diluted 1/200 in PBS plus 1% FCS). Detection with anti-mouse rhodamin labeled secondary antibody. Unstained areas represent nucleoli and heterochromatin as judged from DAPI overlay of the same cell in the right image. This image was kindly supplied as part of the review submitted by Christian Schoefer.
Product References
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