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Product name
Anti-Histone H4 (symmetric di methyl R3) antibody - ChIP Grade
See all Histone H4 products (10) ...
Description
Rabbit polyclonal to Histone H4 (symmetric di methyl R3) - ChIP Grade
Specificity
Current batches recognise symmetric di-methylated H4 but do not cross react with di-methylated R3 histone H2A (see image below). Previous batches have been reported to recognise both symmetric di-methylated H4 and di-methylated R3 histone H2A. This specificity was demonstrated by Ancelin et al., 2006
Tested applications
IHC-P, ChIP, ICC/IF, WBmore details
Cross reactivity
Reacts with
Mouse, Cow, Human, Saccharomyces cerevisiae, Xenopus laevis, Arabidopsis thaliana, Tobacco
Predicted to work with
Rat, Pig, Rainbow Trout, Corn
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 Histone H4, symmetric di methylated at R3.
(Peptide available as ab147 91.)
Positive control
Calf Thymus Histone Preparation; Hela whole cell extract
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
pH: 7.40
Preservative: 0.2% Sodium azide
Constituent: PBS
Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
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Immunocytochemistry/ Immunofluorescence - Histone H2A + H4 (symmetric di methyl R3) antibody - ChIP Grade (ab5823)
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Western blot - Histone H4 (symmetric di methyl R3) antibody - ChIP Grade (ab5823)
(enlarge)
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ChIP - Histone H4 (symmetric di methyl R3) antibody - ChIP Grade (ab5823)
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Applications
Show applications key
Our Abpromise guarantee covers the use of ab5823 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application notes
ChIP: Use 2-4µg for 25µg of chromatin
ICC/IF: 1/2000.
IHC-P: 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB: Use at an assay dependent dilution. Detects a band of approximately 16 kDa (predicted molecular weight: 11.5 kDa). Can be blocked with Histone H4 peptide - symmetric di methyl R3 (ab14791).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Target
Relevance
Histone proteins H3, H4, H2A, and H2B function as building blocks to package eukaryotic DNA into repeating nucleosome units that are folded in higher order chromatin fibers. The nucleosome is composed of an octamer containing a H3/H4 tetramer and two H2A/H2B dimers, surrounded by approximately 146 base pairs of DNA. A diverse and elaborate array of post-translational modifications including acetylation, phosphorylation, methylation, ubiquitination, and ADP-ribosylation occurs on the N-terminal tail domains of histones. Methylation of position-specific lysine residues in histone N termini is a central modification for regulating epigenetic transitions in chromatin. Each methylatable lysine residue can exist in a mono, di, or tri methylated state. Arginine resdiues can also by mono or di methylated.
Cellular localization
Nuclear
Alternative names
- H2a antibody
- H2AFR antibody
- H2AR3me2s antibody
see all
Database links
- Entrez Gene: 8370 Human
- Entrez Gene: 121504 Human
- Entrez Gene: 320332 Mouse
- GenBank: NM_003548 Human
- Omim: 142750 Human
- SwissProt: P59259 Arabidopsis thaliana
- SwissProt: P02304 Human
- SwissProt: P62805 Human
see all
Anti-Histone H4 (symmetric di methyl R3) antibody - ChIP Grade images:
Immunocytochemistry/ Immunofluorescence - Histone H2A + H4 (symmetric di methyl R3) antibody - ChIP Grade (ab5823)
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.
This image is courtesy of an Abreview submitted by Dr Kirk McManus
Western blot - Histone H4 (symmetric di methyl R3) antibody - ChIP Grade (ab5823)
All lanes : Anti-Histone H4 (symmetric di methyl R3) antibody - ChIP Grade (ab5823) at 1 µg/ml
Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate
Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H4 peptide - unmodified (ab15821) at 0.5 µg/ml
Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate with
Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate with
Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate with
Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate with
Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate with
Lysates/proteins at 0.5 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 11.5 kDa
Observed band size : 14 kDa (why is the actual band size different from the predicted?)
ChIP - Histone H4 (symmetric di methyl R3) antibody - ChIP Grade (ab5823)
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.
References for Anti-Histone H4 (symmetric di methyl R3) antibody - ChIP Grade (ab5823)
This product has been referenced in:
- Diring J et al. A Cytoplasmic Negative Regulator Isoform of ATF7 Impairs ATF7 and ATF2 Phosphorylation and Transcriptional Activity. PLoS One 6:e23351 (2011). WB; Tobacco.Read more (PubMed: 21858082) »
- Ahmad A et al. Characterization of the PRMT Gene Family in Rice Reveals Conservation of Arginine Methylation. PLoS One 6:e22664 (2011). WB.Read more (PubMed: 21853042) »
See all 19 publications for this product
Publishing research using ab5823? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"