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Products:Epigenetics and Nuclear Signaling >> Histones >> H3 >> Methylated
Overview
Product name
Anti-Histone H3 (mono methyl K79) antibody - ChIP Grade
See all Histone H3 products (41) ...
Description
Rabbit polyclonal to Histone H3 (mono methyl K79) - ChIP Grade
Specificity
Reacts with Mono-methyl K79 of histone H3. Slight cross-reactivity to di-methyl K79.
Tested applications
ChIP, IF, ICC, IP, Immunodiffusion, ICC/IF, WBmore details
Cross reactivity
Reacts with
Mouse, Rat, Human
Predicted to work with
Saccharomyces cerevisiae, a wide range of other species, all Mammals
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 50 to the C-terminus of Human Histone H3, mono methylated at K79.
(Peptide available as ab4555.)
Positive control
This antibody gave a positive signal in the following lysates: calf thymus histone; HeLa; NIH3T3; rat testis tissue.
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Epigenetics and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Epigenetics and Nuclear Signaling >> Histones >> H3 >> Methylated
Applications
Show applications key
Our Abpromise guarantee covers the use of ab2886 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ChIP: Use 2µg for 106 cells.
IF: Use at an assay dependent dilution.
ICC: Use at an assay dependent dilution.
IP: Use at an assay dependent concentration.
ID: Use at an assay dependent dilution.
ICC/IF: Use at an assay dependent dilution. (PubMed: 18787701)
WB: 1/1 - 1/500.Detects a band of approximately 17 kDa.Can be blocked with Histone H3 peptide - mono methyl K79 (ab4555).
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
- FLJ92264 antibody
- H3 histone antibody
see all
Database links
- Entrez Gene: 8350 Human
- Entrez Gene: 8351 Human
- Entrez Gene: 8352 Human
- Entrez Gene: 8353 Human
- Entrez Gene: 8354 Human
- Entrez Gene: 8355 Human
- Entrez Gene: 8356 Human
- Entrez Gene: 8357 Human
see all
Anti-Histone H3 (mono methyl K79) antibody - ChIP Grade images:
Western blot
All lanes : Anti-Histone H3 (mono methyl K79) antibody - ChIP Grade (ab2886) at 1/500 dilution
Lane 1 : Calf thymus histone lysate
Lane 2 : Calf thymus histone lysate with Histone H3 peptide - mono methyl K79 (ab4555) at 1 µg/ml
Lane 3 : Calf thymus histone lysate with Histone H3 peptide - di methyl K79 (ab4556) at 1 µg/ml
Lane 4 : Calf thymus histone lysate with Histone H3 peptide - tri methyl K79 (ab4557) at 1 µg/ml
Lane 5 : Calf thymus histone lysate with Histone H4 peptide - di methyl K79 (ab4560) at 1 µg/ml
Lane 6 : Calf thymus histone lysate with Histone H3 peptide - mono methyl K9 (ab1771) at 1 µg/ml
Lane 7 : Calf thymus histone lysate with Unmodified K79 of Histone H3 peptide (ab4264) at 1 µg/ml
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 1/5000 dilution
Predicted band size : 15.4 kDa
Observed band size : 17 kDa (why is the actual band size different from the predicted?)
Additional bands at : 14 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 5 minutes
ab2886 specifically detects Histone H3 mono methyl K79. As can be seen in Lane 2, the addition of the blocking peptide (ab4555) quenches the activity of the antibody.
ChIP - Histone H3 (mono methyl K79) antibody - ChIP Grade (ab2886)
Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 µg of chromatin, 2 µg of ab2886 (blue), and 20 µl of protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR. Primers and probes are located in the first kb of the transcribed region.
Immunocytochemistry/ Immunofluorescence - Histone H3 (mono methyl K79) antibody - ChIP Grade (ab2886)
This image was kindly supplied as part of the review submitted by Dr Geza Fejes-Toth.
ChIP - Histone H3 (mono methyl K79) antibody - ChIP Grade (ab2886)
Chromatin was prepared from whole cell lysate of normal rat liver and liver cancer cells. The cross-linking (X-ChiP) technique was used, crosslinking was performed for 15 minutes in formaldehyde. 5 µg of the primary antibody was used in 1/100 dilution and it was incubated with the sample for 16 hours at 4°C in a commercially available ChIP dilution buffer. The immunoprecipitated DNA was quantified by real time PCR. ChIP results show that the Histone H3 (mono methyl K79) and GAPDH genes are expressed in higher levels in liver cancer cells than in normal liver cells.
This image is a courtesy of Anonymous Abreview
Western blot - Histone H3 (mono methyl K79) antibody - ChIP Grade (ab2886)
All lanes : Anti-Histone H3 (mono methyl K79) antibody - ChIP Grade (ab2886) at 1/300 dilution
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 : Testis (Rat) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 15.4 kDa
Observed band size : 18 kDa (why is the actual band size different from the predicted?)
Exposure time : 4 minutes
Immunoprecipitation - Anti-Histone H3 (mono methyl K79) antibody - ChIP Grade (ab2886)
Histone H3 (mono methyl K79) was immunoprecipitated using 0.5mg Hela whole cell extract, 5ug of Rabbit polyclonal to Histone H3 (mono methyl K79) and 50µl of protein G magnetic beads (lane 1). The antibody was incubated with the Protein G beads for 10min under agitation. No antibody was added to the control (lane 2). Hela whole cell extract diluted in RIPA buffer was added to each sample and incubated for 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab2886. Secondary: Clean blot (HRP conjugate) at 1/1000 dilution. Band: 18kDa: Histone H3 (mono methyl K79).
References for Anti-Histone H3 (mono methyl K79) antibody - ChIP Grade (ab2886)
This product has been referenced in:
- Vernimmen Det al. Polycomb eviction as a new distant enhancer function. Genes Dev 25:1583-8 (2011). ChIP; Mouse.Read more (PubMed: 21828268) ��
- Tran BNet al. Novel Histone H3 Binding Protein ORF158L from the Singapore Grouper Iridovirus. J Virol 85:9159-66 (2011). WB.Read more (PubMed: 21715494) »
See all 12 publications for this product
Publishing research using ab2886? 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"