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

• Supplier: Abcam
• Catalog: ab11946
• Prices: $398.00
• Sizes: 100 µg
• Host: Goat
• Clonality: Polyclonal
• Isotype: IgG
• Applications: IHC-P ChIP ICC/IF ICC/IF WB
• Species Reactivities: Mouse, Rat, Bovine, Human, Yeast, Arabidopsis thaliana, C. elegans, Chicken, Xenopus, Drosophila, S. pombe, Zebrafish, Broad species, Tobacco, Chlamydomonas reinhardtii, Aspergillus, Neurospora crassa
• Antigen: Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3 di methylated at K4
• Description: Goat Polyclonal
• 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, 2µg of  ab11946 (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.

All lanes : Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab11946) at 1 µg/mlLane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell LysateLysates/proteins at 15 µg per lane.SecondaryRabbit polyclonal to Goat IgG - H&L - Pre-Adsorbed (HRP) (ab65486) at 1/3000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

Lane 1 : MarkerLanes 2 - 12 : Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab11946) at 1 µg/mlLane 1 : As aboveLane 2 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µgLane 3 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg with Human Histone H3 (unmodified ) peptide (ab7228) at 0.75 µg/mlLane 4 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg with Human Histone H3 (mono methyl K4) peptide (ab1340) at 0.75 µg/mlLane 5 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg with Human Histone H3 (di methyl K4) peptide (ab7768) at 0.75 µg/mlLane 6 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg with Human Histone H3 (tri methyl K4) peptide (ab1342) at 0.75 µg/mlLane 7 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg with Human Histone H3 (mono methyl K9) peptide (ab1771) at 0.75 µg/mlLane 8 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg with Human Histone H3

Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab11946) at 1 µg/ml + PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µgSecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutionPerformed under reducing conditions.

ab11946 (1/500) staining Histone H3 (di methyl K4) in assynchronous HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilized with 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.See Abreview

IHC image of Histone H3 staining in human normal cervix formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. 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 ab11946, 1µg/ml, for 15 mins at room temperature. A donkey anti-goat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

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