Anti-Histone H4 (symmetric di methyl R3) antibody - ChIP Grade

• Supplier: Abcam
• Catalog: ab5823
• Prices: $403.00
• Sizes: 50 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: IHC-P Array ChIP ICC/IF ICC/IF WB
• Species Reactivities: Mouse, Bovine, Human, Yeast, Xenopus, Arabidopsis thaliana, Drosophila, Tobacco, Rat, Pig, Rainbow trout, Corn
• Antigen: Synthetic peptide conjugated to KLH derived from within residues 1 - 100 Histone H4, symmetric di methylated at R3
• Blocking Peptide: Human Histone H4 (symmetric di methyl R3) peptide
• Description: Rabbit Polyclonal
• Gene: His4:CG31611
• Conjugate: Unconjugated

Product images

Anti-Histone H4 (symmetric di methyl R3) antibody - ChIP Grade (ab5823) at 1 µg + Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µgSecondaryAnti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866) at 1/50000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

All batches of ab5823 are tested in Peptide Array against peptides to different Histone H4 and H2A 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 H4 - symmetric di methyl R3 peptide (ab14791), indicating that this antibody specifically recognises the Histone H4 - symmetric di methyl R3 modification.ab14791 - Histone H4 - symmetric di methyl R3ab17416 - Histone H4 asymmetric di methyl R3ab15821 - Histone H4 - unmodifiedab22399 - Histone H2A symmetric di methyl R3ab22398 - Histone H2A asymmetric di methyl R3ab13186 - Histone H2A - unmodified

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  ab5823 (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.

IHC image of ab5823 staining Histone H4 (symmetric di methyl R3) in human kidney formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. 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 ab5823, 5µg/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. No primary antibody was used in the negative control (shown on the inset).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.

Interphase HeLa cell immunofluorescently labelled with anti-dimethylArg3 (H2A + H4) antibody (ab5823, 1/2000) and a polyclonal goat anti-Rabbit secondary antibody conjugated to Cy3 (1/200). ab5823 shows a strong nuclear focal staining pattern. For more information please see Abreview.See Abreview

ab5823 staining Histone H4 (symmetric di methyl R3) in human differentiated haematopoietic stem cells by Flow Cytometry. Cells were fixed with paraformaldehyde and permeabilized with permeabilization buffer. The sample was incubated with the primary antibody (1/400) for 12 hours at 4°C. An Allophycocyanin-conjugated goat monoclonal anti-rabbit IgG (1/1000) was used as the secondary antibody.Gating Strategy: Isotype negative control (white).See Abreview

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