Anti-Histone H3 (methylated) antibody (ab8895)
Overview
- Product nameAnti-Histone H3 (methylated) antibodySee all Histone H3 (methylated) primary antibodies ...
- DescriptionRabbit polyclonal to Histone H3 (methylated)
- SpecificitySpecific for mono-methylated Lysine 4 of histone H3. Does not recognise di- or tri-methyl Lysine 4 nor methylation at Lysine 9.
- Tested applicationsIP, Flow Cyt, Electron Microscopy, ICC/IF, ICC, ChIP, WB, CHIPseq, IHC-P, ChIP/Chip more details
- Species reactivityReacts with: Mouse, Human, Pig, Tetrahymena sp., Xenopus laevis, Plasmodium falciparum
Predicted to work with: Cow, Indian Muntjac, Plants, all Mammals - Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, mono methylated at K4.
(Peptide available as ab1340.)
- Positive controlCalf Thymus Histone Preparation; Hela whole cell extract
Properties
- FormLiquid
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferpH: 7.40
Preservative: 0.02% 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 information loading... - PurityImmunogen affinity purified
- Clonality Polyclonal
- IsotypeIgG
- Research Areas
Applications
Our Abpromise guarantee covers the use of ab8895 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| IP | IP: Use at an assay dependent concentration. |
| Flow Cyt | Flow Cyt: 1/100. |
| Electron Microscopy | EM: Use at an assay dependent dilution. PubMed: 20543957 |
| ICC/IF | ICC/IF: Use at an assay dependent dilution. |
| ICC | ICC: Use at an assay dependent concentration. |
| ChIP | ChIP: Use 2 µg for 25 µg of chromatin. |
| WB | WB: 1/500. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Histone H3 peptide - mono methyl K4 (ab1340). |
| CHIPseq | CHIPseq: Use at an assay dependent concentration. PubMed: 22196736 |
| IHC-P | IHC-P: Use at an assay dependent dilution. |
| ChIP/Chip | ChIP/Chip: Use at an assay dependent dilution. |
Target
- FunctionCore 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 similaritiesBelongs to the histone H3 family.
- Developmental stageExpressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
- Post-translational
modificationsAcetylation 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 localizationNucleus. Chromosome.
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Database links
- 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
- Entrez Gene: 8356 Human
- Entrez Gene: 8357 Human
- Entrez Gene: 8358 Human
- Entrez Gene: 8968 Human
- Entrez Gene: 319152 Mouse
- Entrez Gene: 319153 Mouse
- Entrez Gene: 360198 Mouse
- Entrez Gene: 97908 Mouse
- Omim: 601128 Human
- SwissProt: P68432 Cow
- SwissProt: Q93081 Human
- SwissProt: Q16695 Human
- SwissProt: P68431 Human
- SwissProt: P68433 Mouse
- Unigene: 132854 Human
- Unigene: 247813 Human
- Unigene: 247814 Human
- Unigene: 248176 Human
- Unigene: 443021 Human
- Unigene: 484990 Human
- Unigene: 532144 Human
- Unigene: 533292 Human
- Unigene: 546315 Human
- Unigene: 586261 Human
- Unigene: 591778 Human
- Unigene: 221301 Mouse
- Unigene: 261657 Mouse
- Unigene: 377874 Mouse
- Unigene: 390558 Mouse
- Unigene: 397328 Mouse
see all
Target information above from: UniProt accession
P68431
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010)
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Alternative names
- FLJ92264 antibodyH3 histone antibodyH3 histone antibody
- H3 histone family, member A antibodyH3/a antibodyH3/b antibodyH3/c antibodyH3/d antibodyh3/f antibodyH3/h antibodyH3/i antibodyH3/j antibodyH3/k antibodyH3/l antibodyH31_HUMAN antibodyH3F1K antibodyH3F3 antibodyH3F3 antibodyH3FA antibodyH3FB antibodyH3FC antibodyH3FD antibodyH3FF antibodyH3FH antibodyH3FI antibodyH3FJ antibodyH3FK antibodyH3FL antibodyHIST1H3A antibodyHIST1H3B antibodyHIST1H3C antibodyHIST1H3D antibodyHIST1H3E antibodyHIST1H3F antibodyHIST1H3G antibodyHIST1H3H antibodyHIST1H3I antibodyHIST1H3J antibodyHIST3H3 antibodyHIST3H3 antibodyHistone 1, H3a antibodyHistone cluster 1, H3a antibodyHistone cluster 1, H3b antibodyHistone cluster 1, H3c antibodyHistone cluster 1, H3d antibodyHistone cluster 1, H3e antibodyHistone cluster 1, H3f antibodyHistone cluster 1, H3g antibodyHistone cluster 1, H3i antibodyHistone cluster 1, H3j antibodyHistone H 3 antibodyHistone H3.1 antibodyHistone H3.1 antibodyHistone H3/a antibodyHistone H3/b antibodyHistone H3/c antibodyHistone H3/d antibodyHistone H3/f antibodyHistone H3/h antibodyHistone H3/i antibodyHistone H3/j antibodyHistone H3/k antibodyHistone H3/l antibody
see all
Anti-Histone H3 (methylated) antibody images
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Western blotAll lanes : Anti-Histone H3 (methylated) antibody (ab8895) at 1/500 dilution
Lane 1 : Calf thymus histone lysate
Lane 2 : Calf thymus histone lysate withHistone H3 peptide - mono methyl K4 (ab1340) at 1 µg/ml
Lane 3 : Calf thymus histone lysate withHistone H3 peptide - di methyl K4 (ab7768) at 1 µg/ml
Lane 4 : Calf thymus histone lysate withHistone H3 peptide - tri methyl K4 (ab1342) at 1 µg/ml
Lane 5 : Calf thymus histone lysate withHistone H3 peptide - mono methyl K9 (ab1771) at 1 µg/ml
Lane 6 : Calf thymus histone lysate withHistone H3 peptide - mono methyl K27 (ab1780) at 1 µg/ml
Lane 7 : Calf thymus histone lysate withHistone H3 peptide - unmodified (ab2903) at 1 µg/ml
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size : 15 kDa
Observed band size : 18 kDa (why is the actual band size different from the predicted?)
Exposure time : 2 minutes -
ChIP - Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895)Chromatin was prepared from U2OS 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 ab8895 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified on the GAPDH and ALDOA (active) and MYO-D (inactive) promoters and over the y-Actin gene (active). Schematic diagram of the y-Actin gene is shown on the top of the figure. Black boxes represent exons and thin lines represent introns. PCR products are depicted as bars under the gene.
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Electron Microscopy - Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895)Image from Kizilyaprak C et al, PLoS One. 2010 Jun 9;5(6):e11039, Fig 6.A: Enlarged region showing the distribution of the H3K4me1 labelling around and within the EC cavities of a murine rod photoreceptor.B: Schematic representation of the labelling for the H3K4me1 mark. The bar represents 260 nm in A and 1 µm in B. -
Dot Blot - Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895)Image courtesy of an anonymous Abreview.ab8895 used in Dot Blot at a 1/1000 dilution, 18 hours at 4°C.The dot blot was done using 0.1ug of peptide and screened multiple histone tail modifications to ensure no cross reactivity. -
Immunocytochemistry/ Immunofluorescence - Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895)Kirk McManus in the lab of Michael Hendzel, Univeristy of AlbertaIndian muntjac fibroblast cells - interphase (top left) and prophase (top right and below), stained with:
Mono Methyl K4 antibody, ab8895, (green)
DAPI: red, top left and right; blue, below
Phospho Ser 10 antibody: red (below) and blue (top right)
The perinuclear and perinucleolar heterochromatin domains do not contain Mono Methyl K4. The Mono Methyl K4, rather, is distributed as small nuclear foci primarily found between DAPI-intense regions of the nucleus. -
Flow Cytometry - Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895)This image is courtesy of an Abreview submitted by Prof Albrecht Müllerab8895 staining mouse embryonic stem cells by flow cytometry. The ES cell colonies were trypsinized and the cells permeabilized before being stained with ab8895 (0.25ug/1.5 x 105 cells). A PE conjugated goat anti-rabbit antibody was used as the secondary.
Protocols
References for Anti-Histone H3 (methylated) antibody (ab8895)
This product has been referenced in:
- Tropberger P et al. Regulation of transcription through acetylation of H3K122 on the lateral surface of the histone octamer. Cell 152:859-72 (2013). WB ; Human . Read more (PubMed: 23415232) »
- Choe MK et al. Functional elements demarcated by histone modifications in breast cancer cells. Biochem Biophys Res Commun 418:475-82 (2012). IP . Read more (PubMed: 22285863) »
