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Products:Epigenetics and Nuclear Signaling >> Histones >> H3 >> Unmodified
Overview
Product name
Anti-Histone H3 antibody - ChIP Grade
See all Histone H3 products (17) ...
Description
Rabbit polyclonal to Histone H3 - ChIP Grade
Tested applications
CHIPseq, Dot Blot, Flow Cyt, IHC-P, ICC/IF, ChIP, IP, WB, IF, ChIP/Chip, ICCmore details
Cross reactivity
Reacts 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
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.)
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
pH: 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
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 >> Unmodified
Applications
Show applications key
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.
CHIPseq: Use at an assay dependent dilution.
Dot Blot
Dot: 1/10,000Dot: 1/10,000
Flow Cyt: Use at an assay dependent dilution.
IHC-P: 1/100Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF: Use at an assay dependent dilution.
ChIP: Use 2µg for 106 cells.
IP: Use at an assay dependent dilution.
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).
IF
IF: 1/500IF: 1/500
ChIP/Chip: Use at an assay dependent dilution.
ICC: Use at an assay dependent dilution.
Target
Function
Variant 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 similarities
Belongs to the histone H3 family.
Developmental stage
Expressed throughout the cell cycle independently of DNA synthesis.
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.
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 localization
Nucleus. Chromosome.
Target information above from: UniProt accessionP84243
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- H3 histone antibody
- H3 histone family member E pseudogene antibody
- H3 histone family member E pseudogene antibody
see all
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: 15079 Mouse
- Entrez Gene: 15080 Mouse
see all
Anti-Histone H3 antibody - ChIP Grade images:
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 with Histone H3 peptide (ab12149) at 1 µg/ml
Lane 5 : Jurkat Whole Cell Lysate with Histone H3 peptide (ab12149) at 1 µg/ml
Lane 6 : 293 Whole Cell Lysate with Histone 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.
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 antibody - ChIP Grade (ab1791)
Hela 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).
Kirk McManus, University of British Columbia
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)
ab1791 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.
This image is courtesy of an Abreview submitted by Prof Albrecht Müller
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H3 antibody - ChIP Grade (ab1791)
ab1791 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.
This image is courtesy of an anonymous Abreview
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?)
ab1791 is tested in western blot on a range of species. We recommend loading higher amounts of protein (20-30ug) to increase the signal in yeast lysates
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)
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.
Image courtesy of an anonymous Abreview.
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 seconds
Image courtesy of Dr Roman Hudec by Abreview.
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.
Western blot - Histone H3 antibody - ChIP Grade (ab1791)
Predicted band size : 15 kDa
John E. Mueller and J. Ruth German (Mary Bryk lab)
References for Anti-Histone H3 antibody - ChIP Grade (ab1791)
This product has been referenced in:
- Xu Cet al. Chemical probes identify a role for histone deacetylase 3 in Friedreich's ataxia gene silencing. Chem Biol 16:980-9 (2009).Read more (PubMed: 19778726) »
- Carrozza MJet al. Histone H3 methylation by Set2 directs deacetylation of coding regions by Rpd3S to suppress spurious intragenic transcription. Cell 123:581-92 (2005).Read more (PubMed: 16286007) »
See all 300 publications for this product
Publishing research using ab1791? 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"