Anti-Histone H3 antibody - ChIP Grade (ab1791)
Overview
- Product nameAnti-Histone H3 antibody - ChIP GradeSee all Histone H3 primary antibodies ...
- DescriptionRabbit polyclonal to Histone H3 - ChIP Grade
- Tested applicationsIHC-Fr, CHIPseq, Dot Blot, Flow Cyt, IHC-P, Electron Microscopy, ICC/IF, ChIP, IP, WB, ChIP/Chip, IHC - Wholemount, ICC more details
- Species reactivityReacts with: Mouse, Rat, Chicken, Dog, Human, Saccharomyces cerevisiae, Xenopus laevis, Arabidopsis thaliana, Caenorhabditis elegans, Fruit fly (Drosophila melanogaster), Indian Muntjac, Schizosaccharomyces pombe, Zebrafish, Trypanosoma cruzi, Neurospora crassa , Toxoplasma gondii, Rice, Other, Cyanidioschyzon merolae
Predicted to work with: a wide range of other species, all Mammals - Immunogen
Synthetic peptide conjugated to KLH derived from within residues 100 to the C-terminus of Human Histone H3.
(Peptide available as ab12149.)
- Positive controlHeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
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 ab1791 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| IHC-Fr | IHC-Fr: Use at an assay dependent concentration. |
| CHIPseq | CHIPseq: Use at an assay dependent dilution. |
| Dot Blot | Dot: 1/10000. |
| Flow Cyt | Flow Cyt: Use at an assay dependent dilution. |
| IHC-P | IHC-P: 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
| Electron Microscopy | EM: 1/50. Customer feedback |
| ICC/IF | ICC/IF: Use at an assay dependent dilution. |
| ChIP | ChIP: Use 2µg for 106 cells. |
| IP | IP: Use at an assay dependent dilution. |
| WB | WB: 1/1000 - 1/5000. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Histone H3 peptide (ab12149). |
| ChIP/Chip | ChIP/Chip: Use at an assay dependent dilution. |
| IHC - Wholemount | IHC - Wmt: Use at an assay dependent concentration. PubMed: 22219645 |
| ICC | ICC: Use at an assay dependent dilution. |
Target
- FunctionVariant histone H3 which replaces conventional H3 in a wide range of nucleosomes in active genes. Constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. Deposited at sites of nucleosomal displacement throughout transcribed genes, suggesting that it represents an epigenetic imprint of transcriptionally active chromatin. 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 throughout the cell cycle independently of DNA synthesis.
- 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.
Specifically enriched in modifications associated with active chromatin such as methylation at Lys-5 (H3K4me), Lys-37 and Lys-80. 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), which are linked to gene repression, are underrepresented. 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. Phosphorylation on Ser-32 (H3S31ph) is specific to regions bordering centromeres in metaphase chromosomes.
Ubiquitinated. Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination. - Cellular localizationNucleus. Chromosome.
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Database links
- Entrez Gene: 176359 Caenorhabditis elegans
- Entrez Gene: 31848 Fruit fly (Drosophila melanogaster)
- Entrez Gene: 33736 Fruit fly (Drosophila melanogaster)
- Entrez Gene: 3020 Human
- Entrez Gene: 3021 Human
- Entrez Gene: 15078 Mouse
- Entrez Gene: 15081 Mouse
- Entrez Gene: 100361558 Rat
- Entrez Gene: 100365096 Rat
- Entrez Gene: 117056 Rat
- Entrez Gene: 379062 Xenopus laevis
- Entrez Gene: 379757 Xenopus laevis
- Entrez Gene: 399418 Xenopus laevis
- Entrez Gene: 432115 Xenopus laevis
- Entrez Gene: 444503 Xenopus laevis
- Entrez Gene: 100331798 Zebrafish
- Entrez Gene: 336231 Zebrafish
- Entrez Gene: 394076 Zebrafish
- Entrez Gene: 406269 Zebrafish
- Entrez Gene: 550262 Zebrafish
- Omim: 601128 Human
- SwissProt: P59226 Arabidopsis thaliana
- SwissProt: Q10453 Caenorhabditis elegans
- SwissProt: P08898 Caenorhabditis elegans
- SwissProt: P02299 Fruit fly (Drosophila melanogaster)
- SwissProt: P84249 Fruit fly (Drosophila melanogaster)
- SwissProt: P84243 Human
- SwissProt: P84244 Mouse
- SwissProt: P07041 Neurospora crassa
- SwissProt: P84245 Rat
- SwissProt: Q6PI79 Xenopus laevis
- SwissProt: Q6PI20 Zebrafish
- Unigene: 2931 Fruit fly (Drosophila melanogaster)
- Unigene: 35099 Fruit fly (Drosophila melanogaster)
- Unigene: 7418 Fruit fly (Drosophila melanogaster)
- Unigene: 180877 Human
- Unigene: 533624 Human
- Unigene: 726012 Human
- Unigene: 138832 Mouse
- Unigene: 18516 Mouse
- Unigene: 315189 Mouse
- Unigene: 316825 Mouse
- Unigene: 322735 Mouse
- Unigene: 371563 Mouse
- Unigene: 442502 Mouse
- Unigene: 106155 Rat
- Unigene: 124815 Rat
- Unigene: 198918 Rat
- Unigene: 29857 Rat
- Unigene: 36404 Xenopus laevis
- Unigene: 39769 Xenopus laevis
- Unigene: 4043 Xenopus laevis
- Unigene: 55418 Xenopus laevis
- Unigene: 77438 Xenopus laevis
- Unigene: 132208 Zebrafish
- Unigene: 42924 Zebrafish
- Unigene: 75577 Zebrafish
- Unigene: 75603 Zebrafish
- Unigene: 77073 Zebrafish
see all
Target information above from: UniProt accession
P84243
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010)
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Alternative names
- H3.3B antibody H3F3A antibodyH3 histone antibody
- H3 histone family member E pseudogene antibodyH3 histone family member E pseudogene antibodyH3.3A antibodyH33_HUMAN antibodyH3F3 antibodyH3F3 antibodyH3f3b antibodyHIST3H3 antibodyHIST3H3 antibodyHistone H3 3 pseudogene antibodyHistone H3 3 pseudogene antibodyHistone H3.3 antibodyPP781 antibody
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Anti-Histone H3 antibody - ChIP Grade images
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Western blot - Histone H3 antibody - ChIP Grade (ab1791)All lanes : Anti-Histone H3 antibody - ChIP Grade (ab1791) at 1/1000 dilution
Lane 1 : A431 Whole Cell Lysate
Lane 2 : Jurkat Whole Cell Lysate
Lane 3 : 293 Whole Cell Lysate
Lane 4 : A431 Whole Cell Lysate withHistone H3 peptide (ab12149) at 1 µg/ml
Lane 5 : Jurkat Whole Cell Lysate withHistone H3 peptide (ab12149) at 1 µg/ml
Lane 6 : 293 Whole Cell Lysate withHistone H3 peptide (ab12149) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L (HRP) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 15 kDa
Observed band size : 17 kDa (why is the actual band size different from the predicted?)
Exposure time : 10 seconds -
ChIP - Histone H3 antibody - ChIP Grade (ab1791)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 ab1791 (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 (Taqman approach). Primers and probes are located in the first kb of the transcribed region.
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Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 antibody - ChIP Grade (ab1791)Kirk McManus, University of British ColumbiaHela cells cultured on coverslips were fixed with 4% paraformaldehyde and then stained with ab1791 (green) at a working dilution of 1/500. The DNA stained with DAPI is shown in red. (100x magnification). -
ChIP - Histone H3 antibody - ChIP Grade (ab1791)Chromatin from Xenopus laevis oocytes was prepared according to the Abcam X-ChIP protocol. Oocytes were fixed with formaldehyde for 10 min. The ChIP was performed with 25 mg of chromatin, 3 mg of ab7834 (anti-H3, light blue) and 3 µg of ab1791 (anti-H3, dark blue), and 20 ml of Protein A/G sepharose beads. A non-specific antibody was used as a control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). -
Flow Cytometry - Histone H3 antibody - ChIP Grade (ab1791)This image is courtesy of an Abreview submitted by Prof Albrecht Müllerab1791 staining mouse embryonic stem cells by flow cytometry (gated on all living cells). The cell colonies were trypsinized and incubated with the antibody 1ug/1.5 x 105cells in a permeabilization buffer. A PE conjugated goat anti-rabbit antibody was used as the secondary. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H3 antibody - ChIP Grade (ab1791)This image is courtesy of an anonymous Abreviewab1791 staining rat testes tissue sections by IHC-P. Sections were formaldehyde fixed and subjected to heat mediated antigen retrieval prior to blocking with 5% serum for 30 minutes at 22°C. The primary antibody was diluted 1/400 and incubated with the sample for 30 minutes at 22°C. A HRP-conjugated goat anti-rabbit antibody diluted 1/400 was used as the secondary.
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Western blot - Histone H3 antibody - ChIP Grade (ab1791)All lanes : Anti-Histone H3 antibody - ChIP Grade (ab1791) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 :NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)
Lane 3 : Drosophila embryo nuclear extract (from melanogaster embryos 0-12Hr)
Lane 4 : S.cerevisiae (Y190) Whole Cell Lysate
Lane 5 : S.pombe Whole Cell Lysate
Lysates/proteins at 10 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 : 15 kDa
Observed band size : 17 kDa (why is the actual band size different from the predicted?) -
Immunocytochemistry/ Immunofluorescence - Histone H3 antibody - ChIP Grade (ab1791)ICC/IF image of ab1791 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab1791, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 5% PFA fixed (5 min) Hek293, HepG2 and MCF7 cells at 1µg/ml, and in 4% PFA fixed (10 min) HeLa and Hek293 cells at 1µg/ml. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H3 antibody - ChIP Grade (ab1791)Image courtesy of an anonymous Abreview.ab1791 staining Histone H3 in human benign nevi tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed. Samples were then blocked with 2.5% normal horse serum for 20 minutes followed by incubation with the primary antibody at a 1/200 dilution for 16 hours at 4°C. An undiluted HRP-conjugated rabbit polyclonal was used as the secondary antibody. -
Western blot - Histone H3 antibody - ChIP Grade (ab1791)All lanes : Anti-Histone H3 antibody - ChIP Grade (ab1791) at 1/10000 dilution
Lane 1 : HeLa nuclear lysate
Lane 2 : HeLa nuclear lysate
Lane 3 : HeLa nuclear lysate
Lysates/proteins at 2 µg per lane.
Secondary
HRP conjugated donkey polyclonal at 1/2000 dilution
developed using the ECL technique
Predicted band size : 15 kDa
Exposure time : 5 secondsImage courtesy of Dr Roman Hudec by Abreview.
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Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 antibody - ChIP Grade (ab1791)ICC/IF image of ab1791 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab1791, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. -
Electron Microscopy - Anti-Histone H3 antibody - ChIP Grade (ab1791)Histone H3 immunogold detection in HeLa cells.
Image courtesy of Gerard Pierron, IGR-Villejuif, France.
Ulltra-thin sections of paraformaldehyde-fixed, Lowicryl K4M embedded cells were incubated sequentially with ab1791 antibody and an anti-rabbit antibody coupled to 10 nm gold particles. Notice absence of chromatin within Interchromatin Granules (IG), a reservoir of spliceosomal snRNPs, and high labeling of patches of condensed chromatin close to the nuclear envelope (arrows). Nu: nucleus, Cyt: cytoplasm.Sample : human
Type : HeLa cells
Fixative : either paraformaldehyde 4% or 1.6% glutaraldehyde in 0.1M Millonig’s buffer.
Embedding : in Lowicryl 4KM at -20°C under UV.
Ultrathin-sections deposited on formvar-coated carbonated EM-grids.
Blocking step: 5% BSA for 30 seconds at RT.
Primary antibody: Ab1791 diluted 1/50 in PBS, for 1h at RT.
Secondary antibody: BBI International anti-rabbit antibody coupled to 10 nm gold particlesRating: excellent
Additional comments: A highly specific antibody working similarly on thin-sections of glutaraldehyde or paraformaldehyde-fixed cells. Ideal for testing antigeneicity, for detecting micronuclei, chromatin content of nuclear organelles or bodies…
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Western blot - Histone H3 antibody - ChIP Grade (ab1791)
Predicted band size : 15 kDaJohn E. Mueller and J. Ruth German (Mary Bryk lab)
References for Anti-Histone H3 antibody - ChIP Grade (ab1791)
This product has been referenced in:
- Lau PN & Cheung P Elucidating combinatorial histone modifications and crosstalks by coupling histone-modifying enzyme with biotin ligase activity. Nucleic Acids Res 41:e49 (2013). WB ; Human . Read more (PubMed: 23258705) »
- Keren-Shaul H et al. Pre-mRNA splicing is a determinant of nucleosome organization. PLoS One 8:e53506 (2013). WB, ChIP ; Human . Read more (PubMed: 23326444) »
