Anti-Histone H3 (di methyl K4) antibody - ChIP Grade
| Name | Anti-Histone H3 (di methyl K4) antibody - ChIP Grade |
|---|---|
| Supplier | Abcam |
| Catalog | ab7766 |
| Prices | $403.00 |
| Sizes | 50 µg |
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Applications | ChIP DB WB ChIP ChIP Array IHC-F ICC/IF ICC/IF IHC-P |
| Species Reactivities | Mouse, Human, Pig, Yeast, Drosophila, S. pombe, Plasmodium, Marmoset, Candida, Mammalian |
| Antigen | Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, di methylated at K4 |
| Blocking Peptide | Human Histone H3 (di methyl K4) peptide |
| Description | Rabbit Polyclonal |
| Gene | HIST3H3 |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab7766 (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.
Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766) at 1 µg/ml + Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µgSecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
All batches of ab7766 are tested in Peptide Array against peptides to different Histone H3 modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - di methyl K4 peptide (ab7768), indicating that this antibody specifically recognises the Histone H3 - di methyl K4 modification.ab1340 - Histone H3 - mono methyl K4ab1342 - Histone H3 - tri methyl K4ab1771 - Histone H3 - mono methyl K9ab1772 - Histone H3 - di methyl K9ab1773 - Histone H3 - tri methyl K9ab1780 - Histone H3 - mono methyl K27ab1781 - Histone H3 - di methyl K27ab1782 - Histone H3 - tri methyl K27ab7228 - Histone H3 - unmodifiedab7768 - Histone H3 - di methyl K4
developed using the ECL techniquePerformed under reducing conditions.
Dimethylated lysine 4 (green) is found in several hundred small nuclear foci that do not colocalize with condensed regions of chromatin (DAPI stained, red). The perinuclear and perinucleolar heterochromatin do not stain with this antibody.
ab7766 at 1/200 staining human U2OS (osteosarcoma) cells by ICC/IF. The cells were paraformaldehyde fixed and then stained with the antibody for 1 hour. A Cy2 ® conjugated donkey anti-rabbit antibody was used as the secondary (green). The image shows uniformal staining of the whole nucleus, with several specles found. The insert shows H3-di methyl K4 of tw2 cells only. DAPI nuclear staining is shown in blue.See Abreview
IHC-P image of Histone H3 (di methyl K4) staining with ab7766 on human renal cancer tissue sections. The sections were subjected to heat-mediated antigen retrieval using citrate buffer pH6.0. The sections were then blocked with 5% serum for 20 minutes at 20°C, before incubation with ab7766 (1/500 dilution) for 45 minutes at 20°C. The secondary was a HRP conjugated goat anti-rabbit polyclonal, used at a 1/400 dilution.See Abreview
ICC/IF image of ab7766 stained HeLa cells. The cells were 4% formaldehyde fixed (10 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 (ab7766, 0.1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, a goat anti-rabbit DyLight® 488 (IgG; H+L) 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. This antibody also gave a positive result in 4% formaldehyde fixed (10 min) Hek293, HepG2 and MCF7 cells at 0.1µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 0.1µg/ml.
Product References
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