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Products:Epigenetics and Nuclear Signaling >> Histones >> H3 >> Methylated
Overview
Product name
Anti-Histone H3 (tri methyl K4) antibody
See all Histone H3 products (41) ...
Description
Rabbit polyclonal to Histone H3 (tri methyl K4)
Specificity
ab71998 is specific for Histone H3 tri-methylated at K4. It does not cross react to nonmethylated H3 or mono-, di-methylated Histone H3.
Tested applications
WB, IP, ELISA, ICC/IFmore details
Cross reactivity
Reacts with
Mouse, Rat, Chicken, Cow, Human
Predicted to work with
Xenopus laevis, Arabidopsis thaliana, Fruit fly (Drosophila melanogaster)
Immunogen
A synthetic peptide containing RT[Me3-K]QT in which Me3-K corresponds to trimethyl lysine 4 of human histone H3.
Positive control
3T3 Nuclear Extract and 3T3 cells.
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
Preservative: None
Constituents: PBS, pH 7.2 containing antibody stabilizer
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Purification notes
ab71998 is purified by Site-modification specific Affinity Chromatography.
Clonality
Polyclonal
Isotype
IgG
Research areas
Epigenetics and Nuclear Signaling >> Histones >> H3 >> Methylated
Applications
Show applications key
Our Abpromise guarantee covers the use of ab71998 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: Use a concentration of 0.1 - 1 µg/ml.Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).(IP is recommended before WB. This antibody is not ChIP grade. Results shown in the image indicate a slight enrichment at heterchromatic loci. This suggests the antibody may exhibit some cross-reactivity with histone modifications localized at these regions. We would welcome feedback about this antibodies performance in ChIP.)
IP: Use a concentration of 2 - 5 µg/ml.
ELISA: Use a concentration of 0.01 - 0.1 µg/ml.
ICC/IF: 1/100
Target
Function
Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
Sequence similarities
Belongs to the histone H3 family.
Developmental stage
Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
Post-translational
modifications
Acetylation is generally linked to gene activation. Acetylation on Lys-10 (H3K9ac) impairs methylation at Arg-9 (H3R8me2s). Acetylation on Lys-19 (H3K18ac) and Lys-24 (H3K24ac) favors methylation at Arg-18 (H3R17me).
Citrullination at Arg-9 (H3R8ci) and/or Arg-18 (H3R17ci) by PADI4 impairs methylation and represses transcription.
Asymmetric dimethylation at Arg-18 (H3R17me2a) by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 (H3R8me2s) by PRMT5 is linked to gene repression. Asymmetric dimethylation at Arg-3 (H3R2me2a) by PRMT6 is linked to gene repression and is mutually exclusive with H3 Lys-5 methylation (H3K4me2 and H3K4me3). H3R2me2a is present at the 3' of genes regardless of their transcription state and is enriched on inactive promoters, while it is absent on active promoters.
Methylation at Lys-5 (H3K4me), Lys-37 (H3K36me) and Lys-80 (H3K79me) are linked to gene activation. Methylation at Lys-5 (H3K4me) facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 (H3K79me) is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are linked to gene repression. Methylation at Lys-10 (H3K9me) is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 (H3S10ph) and acetylation of H3 and H4. Methylation at Lys-5 (H3K4me) and Lys-80 (H3K79me) require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are enriched in inactive X chromosome chromatin.
Phosphorylated at Thr-4 (H3T3ph) by GSG2/haspin during prophase and dephosphorylated during anaphase. Phosphorylation at Ser-11 (H3S10ph) by AURKB is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at Ser-11 (H3S10ph) by RPS6KA4 and RPS6KA5 is important during interphase because it enables the transcription of genes following external stimulation, like mitogens, stress, growth factors or UV irradiation and result in the activation of genes, such as c-fos and c-jun. Phosphorylation at Ser-11 (H3S10ph), which is linked to gene activation, prevents methylation at Lys-10 (H3K9me) but facilitates acetylation of H3 and H4. Phosphorylation at Ser-11 (H3S10ph) by AURKB mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. Phosphorylation at Ser-11 (H3S10ph) is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylated at Ser-29 (H3S28ph) by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation at Thr-7 (H3T6ph) by PRKCBB is a specific tag for epigenetic transcriptional activation that prevents demethylation of Lys-5 (H3K4me) by LSD1/KDM1A. At centromeres, specifically phosphorylated at Thr-12 (H3T11ph) from prophase to early anaphase, by DAPK3 and PKN1. Phosphorylation at Thr-12 (H3T11ph) by PKN1 is a specific tag for epigenetic transcriptional activation that promotes demethylation of Lys-10 (H3K9me) by KDM4C/JMJD2C. Phosphorylation at Tyr-42 (H3Y41ph) by JAK2 promotes exclusion of CBX5 (HP1 alpha) from chromatin.
Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination (By similarity). Ubiquitinated by the CUL4-DDB-RBX1 complex in response to ultraviolet irradiation. This may weaken the interaction between histones and DNA and facilitate DNA accessibility to repair proteins.
Cellular localization
Nucleus. Chromosome.
Target information above from: UniProt accessionP68431
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- Histone H3.1 antibody
- Histone H3.1 antibody
- FLJ92264 antibody
see all
Database links
- Entrez Gene: 427887 Chicken
- Entrez Gene: 326601 Cow
- Entrez Gene: 8350 Human
- Entrez Gene: 8351 Human
- Entrez Gene: 8352 Human
- Entrez Gene: 8353 Human
- Entrez Gene: 8354 Human
- Entrez Gene: 8355 Human
see all
Anti-Histone H3 (tri methyl K4) antibody images:
Immunocytochemistry/ Immunofluorescence - Histone H3 (tri methyl K4) antibody (ab71998)
ab71998 at 1/100 dilution staining Histone H3 in 3T3 cells by Immunocytochemistry/ Immunofluorescence. A FITC labeled goat anti Rb IgG was used for visualization under fluorescent microscope.
Western blot - Histone H3 (tri methyl K4) antibody (ab71998)
All lanes : Anti-Histone H3 (tri methyl K4) antibody (ab71998) at 1/1000 dilution
Lane 1 : 3T3 nuclear extract immunoprecipitated
using 4 µg ab71998
Lane 2 : 3T3 nuclear extract immunoprecipitated
using control Rabbit IgG
Predicted band size : 15 kDa
Observed band size : 17 kDa (why is the actual band size different from the predicted?)
Additional bands at : 22 kDa (possible IgG),50 kDa (possible IgG).
For IP 5 mg/ml of cell nuclear extract was incubated with 4 ug of antibody 4oC O/N and pulled down by Protein A beads. Samples were then boiled in SDS-PAGE sample buffer and loaded into 12% SDS-PAGE. Transfer was onto NC membrane and blotted with the same antibody at 1:1000.
ChIP - Histone H3 (tri methyl K4) antibody (ab71998)
Chromatin was prepared from Jurkat 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 ab71998 (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 H3 (tri methyl K4) antibody (ab71998)
ab71998 has not yet been referenced specifically in any publications.
Publishing research using ab71998? 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"