Key features and details
- Rabbit polyclonal to Histone H3 - Nuclear Marker and ChIP Grade
- Suitable for: IHC-P, Electron Microscopy, ChIP, IP, WB
- Reacts with: Mouse, Rat, Chicken, Dog, Human, Saccharomyces cerevisiae, Xenopus laevis, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster, Ferret, Indian muntjac, Schizosaccharomyces pombe, Zebrafish, Silk worm, Dictyostelium discoideum, Rainbow trout, Trypanosoma cruzi, Neurospora crassa, Toxoplasma gondii, Rice, Schistosoma mansoni, Cyanidioschyzon merolae
- Isotype: IgG
製品名Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade
Histone H3 一次抗体 製品一覧
製品の詳細Rabbit polyclonal to Histone H3 - Nuclear Marker and ChIP Grade
アプリケーション適用あり: IHC-P, Electron Microscopy, ChIP, IP, WBmore details
種交差性交差種: Mouse, Rat, Chicken, Dog, Human, Saccharomyces cerevisiae, Xenopus laevis, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster, Ferret, Indian muntjac, Schizosaccharomyces pombe, Zebrafish, Silk worm, Dictyostelium discoideum, Rainbow trout, Trypanosoma cruzi, Neurospora crassa, Toxoplasma gondii, Rice, Schistosoma mansoni, Candida albicans, Cyanidioschyzon merolae
交差が予測される動物種: a wide range of other species, Mammals
- WB: A431, Jurkat, HEK293, HeLa, NIH 3T3, Drosophila embryo nuclear extract, S.cerevisiae (Y190), S.pombe whole cell lysates, mouse skeletal muscle mitochondrial fraction. ICC/IF: HeLa. ChIP: Chromatin from HeLa cells, chromatin from Xenopus laevis oocytes, chromatin from mouse gut cell lysate. ChIPSeq: SNA1 KO MEFs. IHC-Fr: Mouse brain tissue. IHC-P: Rat brain tissue, human infantile fibromatosis tissue, mouse liver tissue. EM: HeLa cells. Flow Cyt: Mouse embryonic stem cells. IP: HeLa cell lysates.
保存方法Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
バッファーPreservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
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...
精製度Immunogen affinity purified
ChIP Related Products
Immunizing Peptide (Blocking)
- Histone H3 Total Quantification Kit (Colorimetric) (ab115091)
- Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
- Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
- Human Histone H3 (tri methyl K4) peptide (ab1342)
- Human Histone H3 (tri methyl K9) peptide (ab1773)
- Human Histone H3 (tri methyl K27) peptide (ab1782)
- Histone H3 Modification Multiplex Assay Kit (Colorimetric) (ab185910)
- Anti-Histone H3 antibody [mAbcam 24834] - Nuclear Loading Control and ChIP Grade (ab24834)
- Anti-TATA binding protein TBP antibody [mAbcam 51841] - ChIP Grade (ab51841)
- Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
- Human Histone H3 (unmodified ) peptide (ab7228)
- Human Histone H3 (di methyl K4) peptide (ab7768)
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.
|IHC-P||1/100 - 1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|Electron Microscopy||1/50. Customer feedback|
|ChIP||Use 2µg for 106 cells.|
|IP||Use a concentration of 5 µg/ml.|
|WB||1/1000 - 1/5000. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 peptide (ab12149).|
機能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.
配列類似性Belongs to the histone H3 family.
発生段階Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
翻訳後修飾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.
- Information by UniProt
- H3 histone family member E pseudogene antibody
- H3 histone family, member A antibody
- H3/A antibody
Histone H3 - ChIP Grade was immunoprecipitated using 0.5mg HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell extract, 5 µg of Rabbit polyclonal to and 50 µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10 minutes, HeLa whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10 minutes under agitation.
Proteins were eluted by addition of 40 µl SDS loading buffer and incubated for 10 minutes at 70°C; 10 µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab1791.
Secondary Antibody: Mouse anti-rabbit HRP light chain (HRP) (ab99697).
Band: 15kDa; Histone H3 - ChIP Grade
Chromatin from Xenopus laevis oocytes was prepared according to the Abcam X-ChIP protocol.
Oocytes were fixed with formaldehyde for 10 minutes. 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).
ab1796 staining Histone H3 in mouse liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).
Tissue was fixed with paraformaldehyde, permeabilized with 0.05% Triton X-100 in PBS for 30 minutes and blocked with 5% BSA for 1 hour; antigen retrieval was by heat mediation in sodium citrate pH 6. Samples were incubated with the primary antibody (1/500 in blocking buffer) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody.
Histone H3 immunogold detection in HeLa cells.
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.
Image courtesy of Gerard Pierron, IGR-Villejuif, France.
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
All lanes : Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791) at 1/1000 dilution
Lane 1 : Mouse skeletal muscle mitochondrial fraction
Lane 2 : Mouse skeletal muscle nuclear fraction
Lysates/proteins at 20 µg per lane.
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/4000 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: 7 minutes
Blocked with 3% milk for 1 hour at 25°C.
Incubated with the primary antibody for 16 hours at 4°C in 3% milk in TBS-tween.
All lanes : Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade (ab1791) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 :
NIH 3T3 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.
All lanes : 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
Rabbit polyclonal to Histone H3 (ab1791) at 1/5000 on S. cerevisiae whole cell lysate (40 ug per lane).
Protein resolved on 15% SDS-PAGE gel. After transfer to PVDF membrane, blots were blocked in 1X PBS, 0.1% Tween-20, and 5% milk. ab1791 was diluted in 5 ml blocking buffer at 1/5000. Blots plus primary antibodies were either incubated overnight at 4°C or at RT for 2 hours. Blots were washed 6X for 10 minutes each in PBS with 0.1% Tween-20 before addition of secondary antibodies. Secondary antibodies were diluted 1/2,000 in blocking buffer and incubated with blots for 2 hours at RT. Secondary blots were washed 4X for 10 minutes each in PBS with 0.1% Tween-20 and 2X for 10 minutes each in PBS.
Chromatin was prepared from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol.
Cells were fixed with formaldehyde for 10 minutes. 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 for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.
The ChIP was performed with chromatin from mouse gut cell lysate and ab1791 at 1/250 dilution.
Negative control: No antibody was used (right bar).
The immunoprecipitated DNA was quantified by real time PCR.
Paraffin-embedded rat brain tissue stained for Histone H3 using ab1791 at 1/8000 dilution in immunohistochemical analysis.
ab1791 staining Histone H3 in human infantile fibromatosis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).
Tissue was fixed with formaldehyde and blocked with 1% FBS/BSA for 3 hours at room temperature; antigen retrieval was by heat mediation in Tris pH 9. Samples were incubated with primary antibody (1/100 in TBS + 1% BSA + 1% FBS) for 16 hours. An un-diluted HRP-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.
ab1791 staining Histone H3 (red) in rat brain tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).
Tissue was fixed with formaldehyde, permeabilized with 0.1% TBS-TritonX and blocked with 10% serum for 1 hour at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with the primary antibody (1/500 in 10% normal goat serum) for 24 hours at 24°C. An Alexa Fluor® 594-conjugated goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.
Green - Nucleus staining.
Red - Histone H3 staining.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab1791 は 3065 報の論文で使用されています。
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