Anti-Histone H3 (tri methyl K79) 抗体 - ChIP Grade (ab2621)
製品の概要
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製品名Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade
Histone H3 一次抗体 製品一覧 -
製品の詳細Rabbit polyclonal to Histone H3 (tri methyl K79) - ChIP Grade
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由来種Rabbit
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特異性Reacts with Tri-methyl K79 of histone H3. Cross-reactivity to di-methyl K79.
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アプリケーション適用あり: ICC/IF, ChIP, WB, ChIP/Chipmore details
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種交差性交差種: Mouse, Rat, Cow, Human, Saccharomyces cerevisiae, Plasmodium falciparum
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免疫原
Synthetic peptide within Human Histone H3 aa 50 to the C-terminus (tri methyl K79) conjugated to Keyhole Limpet Haemocyanin (KLH). The exact sequence is proprietary.
(Peptide available asab4557) -
ポジティブ･コントロール
- WB: Calf thymus histone preparation and HeLa whole cell extract.
製品の特性
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製品の状態Liquid
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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.
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バッファーpH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
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
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ポリ/モノポリクローナル
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アイソタイプIgG
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研究分野
関連製品
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ChIP Related Products
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Compatible Secondaries
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Control Peptide
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Immunizing Peptide (Blocking)
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Isotype control
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Related Products
アプリケーション
Our Abpromise guarantee covers the use of ab2621 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| アプリケーション | Abreviews | 特記事項 |
|---|---|---|
| ICC/IF | Use at an assay dependent concentration. | |
| ChIP | Use 2-4 µg for 6 µg of chromatin. | |
| WB | Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 (tri methyl K79) peptide (ab4557). Abcam recommends using BSA as the blocking agent. |
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| ChIP/Chip | Use at an assay dependent concentration. |
ターゲット情報
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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.
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配列類似性Belongs to the histone H3 family.
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発生段階Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
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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. -
細胞内局在Nucleus. Chromosome.
- Information by UniProt
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参照データベース
- 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
see all -
別名
- H3 histone family member E pseudogene antibody
- H3 histone family, member A antibody
- H3/A antibody
see all
画像
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ChIP - Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)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 ab2621 (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. Primers and probes are located in the first kb of the transcribed region.
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Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)This image is courtesy of an anonymous abreview.Immunocytochemistry/ Immunofluorescence analysis of HEK293 cells labeling Histone H3 (tri methyl K79) with ab2621 at 1/1000 dilution. Cells were fixed with paraformaldehyde and permeabilized with Triton-X. The cells were blocked with 5% BSA for 1 hour at 21°C, followed by incubation with Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621) in 3% BSA in TBST for 16 hours at 4°C. A goat anti-rabbit Alexa Fluro 594 secondary antibody was used at 1/1000 dilution.
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Western blot - Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)All lanes : Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621) at 1 µg/ml
Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate
Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (unmodified ) peptide (ab4558) at 0.5 µg/ml
Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K79) peptide (ab4555) at 0.5 µg/ml
Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate withHuman Histone H3 (di methyl K79) peptide (ab4556) at 0.5 µg/ml
Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate withHuman Histone H3 (tri methyl K79) peptide (ab4557) at 0.5 µg/ml
Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H4 (di methyl K79) peptide (ab4560) at 0.5 µg/ml
Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate withHuman Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µg/ml
Lysates/proteins at 0.5 µg per lane.
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 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 - Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)This image is courtesy of an Abreview submitted by Dr Kirk McManusab2621 (1/400) staining Histone H3 (tri Methyl K79) in human hTERT cells (green). Cells were fixed in methanol, permeabilised with 0.5% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details, or to check results obtained using paraformaldehyde fixation, please refer to abreview. -
ChIP - Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)This image is a courtesy of Anonymous AbreviewChromatin was prepared from whole cell lysate of normal rat liver and liver cancer cells. The cross-linking (X-ChiP) technique was used, crosslinking was performed for 15 minutes in formaldehyde. 5 µg of the primary antibody was used in 1/100 dilution and it was incubated with the sample for 16 hours at 4°C. The immunoprecipitated DNA was quantified by real time PCR.
参考文献
This product has been referenced in:
- Zhu B et al. The protective role of DOT1L in UV-induced melanomagenesis. Nat Commun 9:259 (2018). Read more (PubMed: 29343685) »
- Zheng S et al. DZNep inhibits H3K27me3 deposition and delays retinal degeneration in the rd1 mice. Cell Death Dis 9:310 (2018). Read more (PubMed: 29472543) »
