Anti-Histone H3 (phospho S10) [mAbcam 14955] antibody – ChIP Grade (ab14955)抗组蛋白H3抗体
货号：ab14955
厂家：Abcam公司
规格：100ug
保存温度：-20℃ 和 2～8℃
货期：现货2～5天，期货2-3周
咨询： （重庆） （贵阳）
Anti-Histone H3 (phospho S10) [mAbcam 14955] antibody – ChIP Grade (ab14955)抗组蛋白H3抗体产品信息：
- Product name:Anti-Histone H3 (phospho S10) [mAbcam 14955] antibody – ChIP Grade
- Description
Mouse monoclonal [mAbcam 14955] to Histone H3 (phospho S10) – ChIP Grade
- Specificityab14955 recognises phospho S10 on Histone H3, even when K9 is tri methylated. It recognises a phospho S28 peptide by ELISA, but is not blocked by phospho S28 in WB.
- Tested applicationsIHC-P, ICC/IF, ChIP, IP, IHC-Fr, Flow Cyt, WB
- Species reactivity
Reacts with: Mouse, Rat, Human, Xenopus laevis, Arabidopsis thaliana, Fruit fly (Drosophila melanogaster), Indian Muntjac, African Green Monkey
Predicted to work with: Chicken, Saccharomyces cerevisiae, Caenorhabditis elegans, Schizosaccharomyces pombe, Zebrafish - Immunogen
Phospho S10 specific clone produced using a synthetic peptide derived from residues 1 – 100 of Human Histone H3, phosphorylated at S10 and tri methylated at K9.
Read Abcam's proprietary immunogen policy
Hybridomas were prepared and the resulting clones were positively screened by ELISA against the immunising tri methyl K9 and phospho S10 dimodified peptide. Clones were also positively screened against both tri methyl K9 and phospho S10 peptides. Clones were negatively screened against the unmodified version of the peptides. This clone binds to the tri methyl K9 and phospho S10 dimodified peptide and to the phospho S10 peptide, but not to the tri methyl K9 peptide or to equivalent unmodified Histone H3 peptide.
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: 7.50
Preservative: 0.02% Sodium azide
Constituents: 5.88% Sodium citrate, Tris HCl, 2.9% Sodium chloride
Some batches contain 0.4M arginine -
Concentration 100 µg at 1 mg/ml
- PurityIgG fraction
- Clonality Monoclonal
- Clone numbermAbcam 14955
- IsotypeIgG1
- Research Areas
- Epigenetics and Nuclear Signaling
- Histones
- H3
- Phosphorylated
- Epigenetics and Nuclear Signaling
- Histones
- H3
- Methyl + Phospho
- Epigenetics and Nuclear Signaling
- ChIP'ing antibodies
- ChIP'ing antibodies
Applications
Our Abpromise guarantee covers the use of ab14955 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-P | IHC-P: Use at an assay dependent dilution. |
| ICC/IF | ICC/IF: Use at an assay dependent dilution. |
| ChIP | ChIP: Use at an assay dependent dilution. PubMed: 20864037See Abreview submitted on 5 January 2007. |
| IP | IP: Use at 80 µg/mg of lysate. |
| IHC-Fr | IHC-Fr: Use at an assay dependent concentration. PubMed: 21734301 |
| Flow Cyt | Flow Cyt: 1/1000. |
| WB | WB: Use a concentration of 1 – 5 µg/ml. Detects a band of approximay 17 kDa (predicted molecular weight: 15 kDa). |
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.
-
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
-
Alternative names
- H3 3 like sequence MH921 antibody
- H3 3 like sequence MH921 antibody
- H3 3A antibody
Anti-Histone H3 (phospho S10) [mAbcam 14955] antibody – ChIP Grade images
Western blot – Histone H3 (phospho S10) antibody [mAbcam 14955] – ChIP Grade (ab14955)
All lanes : Anti-Histone H3 (phospho S10) [mAbcam 14955] antibody – ChIP Grade (ab14955) at 1 µg/ml
Lane 1 : Control HeLa Histone Prep 0.5ug
Lane 2 : Colcemid treated HeLa Histone prep 0.5ug
Lane 3 : Control HeLa Histone Prep 0.5ug with Human Histone H3 (tri methyl K9, phospho S10) peptide (ab15644) at 1 µg/ml
Lane 4 : Colcemid treated HeLa Histone prep 0.5ug with Human Histone H3 (tri methyl K9, phospho S10) peptide (ab15644) at 1 µg/ml
Lane 5 : Control HeLa Histone Prep 0.5ug with Human Histone H3 (unmodified ) peptide (ab7228) at 1 µg/ml
Lane 6 : Colcemid treated HeLa Histone prep 0.5ug with Human Histone H3 (unmodified ) peptide (ab7228) at 1 µg/ml
Lane 7 : Control HeLa Histone Prep 0.5ug with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml
Lane 8 : Colcemid treated HeLa Histone prep 0.5ug with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml
Lane 9 : Control HeLa Histone Prep 0.5ug with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml
Lane 10 : Colcemid treated HeLa Histone prep 0.5ug with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml
Secondary
Rabbit polyclonal to Mouse IgG H&L (HRP) at 1/5000 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 (phospho S10) antibody [mAbcam 14955] (ab14955)This image was submitted as part of a review by Kirk McManus
Indian muntjac and HeLa cells were fixed in paraformaldehyde and labelled with ab14955 (1/2000 dilution) for 30 minutes. The above image contains an interphase and a prophase Indian muntjac cells immunofluorescently labelled with anti-PhosS10 (green) and counterstained with DAPI (red).
ELISA – Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)
By ELISA, ab14955 detects:
the singly modified phospho S10 peptide and the dual modified phospho S10 and tri methyl K9 peptide (the orange and grey lines respectively),
less strongly detects the phospho S28 peptide (purple line),
does not detect the equivalent non-modified Histone H3 peptide for S10 or the singly modified tri methyl K9 peptide (the 2 lines at the bottom of the figure).The antibody recognises phospho S28 by ELISA (although a phospho S28 peptide does not block the antibody in Western blotting).
Flow Cytometry – Histone H3 (phospho S10) antibody [mAbcam 14955] – ChIP Grade (ab14955)This image is courtesy of an Abreview submitted by Dr Kirk McManus
ab14955 (1/1000) stainning a population of Human HeLa cells positive for Histone H3 (phospho S10). Cells were trypsizined, pelleted and fixed in ice cold ethanol. Cellular debris was eliminated and the FL2-A/FL2-W was used to eliminate clumping cells. For further experimental details please refer to abreview.
-
Western blot – Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)
Lane 1 – 8 : Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955) at 0.5 ug/ml
Lane 1 : Control HeLa Histone prep at 5 ug
Lane 2 : Colcemid treated HeLa Histone prep at 5 ug
Lane 3 : Control HeLa Histone prep at 5 ug with Histone H3 peptide – unmodified (ab7228) at 1 ug/ml
Lane 4 : Colcemid treated HeLa Histone prep at 5 ug with Histone H3 peptide – unmodified (ab7228) at 1 ug/ml
Lane 5 : Control HeLa Histone prep at 5 ug with Histone H3 peptide – phospho S28 (ab14793) at 1 ug/ml
Lane 6 : Colcemid treated HeLa Histone prep at 5 ug with Histone H3 peptide – phospho S28 (ab14793) at 1 ug/ml
Lane 7 : Control HeLa Histone prep at 5 ug with Histone H3 peptide – phospho S10 (ab11477) at 1 ug/mlThe antibody is not blocked by a phospho S28 peptide by Western Blotting.
Western blot – Anti-Histone H3 (phospho S10) [mAbcam 14955] antibody – ChIP Grade (ab14955)Image courtesy of Richelle Sopko, Harvard University,U.S.A
All lanes : Anti-Histone H3 (phospho S10) [mAbcam 14955] antibody – ChIP Grade (ab14955) at 1/1000 dilution
Lane 1 : Wild type 0-4 hour old fruit fly embryo lysate
Lane 2 : 0-4 hour old fruit fly embryo lysate expressing wee RNAi
Secondary
Anti-mouse IgG, peroxidase-linked at 1/10000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 15 kDa
Exposure time : 5 secondsImage courtesy of Richelle Sopko, Harvard University, U.S.A
Blocking: 10% BSA
wee shRNA embryos (lane 2) should display elevated phospho H3Ser10 levels relative to wild type
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