Anti-Histone H3 (di methyl K9) 抗体 [mAbcam 1220] - ChIP Grade (ab1220)
Key features and details
- Mouse monoclonal [mAbcam 1220] to Histone H3 (di methyl K9) - ChIP Grade
- Suitable for: WB, ELISA, IHC-P, ChIP
- Reacts with: Rat, Chicken, Cow, Human, Xenopus laevis, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe, Corn, Common marmoset, Rice
- Isotype: IgG2a
製品の概要
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製品名
Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade
Histone H3 一次抗体 製品一覧 -
製品の詳細
Mouse monoclonal [mAbcam 1220] to Histone H3 (di methyl K9) - ChIP Grade -
由来種
Mouse -
特異性
By peptide ELISA ab1220 recognizes di methyl K9, but not unmodified K9, mono methyl K9, tri methyl K9, di methyl K27, tri methyl K27, mono methyl K4, di methyl K4 or tri methyl K4. By Western blot ab1220 is blocked by di methyl K9, but not by unmodified K9, mono methyl K9, tri methyl K9, di methyl K27, tri methyl K27, mono methyl K4, di methyl K4 or tri methyl K4. This indicates the specificity of ab1220 for di methyl K9 of Histone H3. -
アプリケーション
適用あり: WB, ELISA, IHC-P, ChIPmore details -
種交差性
交差種: Rat, Chicken, Cow, Human, Xenopus laevis, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe, Corn, Common marmoset, Rice
交差が予測される動物種: Mouse, Sheep, Saccharomyces cerevisiae, Other species -
免疫原
Synthetic peptide corresponding to Human Histone H3 aa 1-100 (di methyl K9) conjugated to keyhole limpet haemocyanin (Cysteine residue). Clones were positively screened by ELISA against the immunising peptide. Clones were negatively screened against both the non-modified equivalent peptide and against a dimethylated K27 peptide.
(Peptide available asab1772) -
ポジティブ・コントロール
- IHC-P: Human kidney tissue; Rat liver tissue; Mouse liver P5 tissue. Flow Cyt: Mouse embryonic stem cells. WB: HeLa whole cell lysate; Calf thymus histone lysate; HEK-293 lysate; Fruit fly embryo tissue lysate. ChIP: AFM ChIP primer pair ab269259; Chromatin prepared from U2OS cells .
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特記事項
Learn about ChIP assay kits, other ChIP antibodies, and more in the ChIP assay guide.
ChIP protocols:
ChIP protocol for cross-linking ChIP (X-ChIP)
Native ChIP protocol
Chromatin preparation from tissues for ChIP
ChIP troubleshooting
ChIP tips and tricks guideThis antibody clone [mAbcam 1220] is manufactured by Abcam.
If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
製品の特性
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製品の状態
Liquid -
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
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading...
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精製度
IgG fraction -
ポリ/モノ
モノクローナル -
クローン名
mAbcam 1220 -
アイソタイプ
IgG2a -
軽鎖の種類
kappa -
研究分野
関連製品
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Alternative Versions
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ChIP Related Products
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Compatible Secondaries
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Corresponding Unmodified Peptide
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Immunizing Peptide (Blocking)
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
- Histone H3 (K9) Methyltransferase Activity Quantification Assay Kit (ab113453)
- Histone H3 (di-methyl K9) Quantification Kit (Colorimetric) (ab115062)
- Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
- Human Histone H3 (tri methyl K4) peptide (ab1342)
- Human Histone H3 (tri methyl K9) peptide (ab1773)
- Human Histone H3 (tri methyl K27) peptide (ab1782)
- Human Histone H3 (unmodified) peptide (ab7228)
- Human Histone H3 (di methyl K4) peptide (ab7768)
アプリケーション
Our Abpromise guarantee covers the use of ab1220 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 |
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WB | Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).Can be blocked with Human Histone H3 (di methyl K9) peptide (ab1772). | |
ELISA | Use a concentration of 1 - 0.00025 µg/ml. when testing with immunogen peptide. | |
IHC-P | Use at an assay dependent concentration. | |
ChIP | Use 2-4 µg for 25 µg of chromatin. Use AFM ChIP primer pair ab269259 as positive control. |
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. -
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. -
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 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)Image from Alarcon V et al., Sci Rep 13(6), Fig 4. Doi: 10.1038/srep25901. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Alarcon V et al investigates the effects of pargyline due to inhibition of LSD1. Levels of LSD1 protein were reduced using siRNA (siLSD1). Huh7 (Human hepatocellular carcinoma) cells were treated with and without siLSD1 and incubated for 24 hours before transfection of HBV genome (A). Western blot analysis shows the reduction of LSD1 after treatment. Covalent post-translational modifications on Histone H3 was determined by ChIP using ab1220 as one of the specific antibodies (C-G). HBV cccDNA and cytoplasmic levels were determined by qPCR (H).
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All lanes : Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
Lane 1 : Calf thymus histone lysate
Lane 2 : Calf thymus histone lysate with Histone H3 peptide - unmodified at 1 µg/ml
Lane 3 : Calf thymus histone lysate withHuman Histone H3 (mono methyl K9) peptide (ab1771) at 1 µg/ml
Lane 4 : Calf thymus histone lysate withHuman Histone H3 (di methyl K9) peptide (ab1772) at 1 µg/ml
Lane 5 : Calf thymus histone lysate withHuman Histone H3 (tri methyl K9) peptide (ab1773) at 1 µg/ml
Lane 6 : Calf thymus histone lysate withHuman Histone H3 (di methyl K4) peptide (ab7768) at 1 µg/ml
Lane 7 : Calf thymus histone lysate withHuman Histone H3 (di methyl K27) peptide (ab1781) at 1 µg/ml
Secondary
All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 17 kDa
Exposure time: 1 minute -
Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25µg of chromatin, 2µg of ab1220 (blue), and 20µl 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.
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ChIP - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)Image from Chen L et al., PLoS One 9(7), Fig 6. Doi: 10.1371/journal.pone.0101040 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Chen L et al investigates role of Trim 28 (Tripartite motif containing 28) in the epithelial to mesenchymal transition which is implicated in cancer metastasis. ChIP was performed using ab1220. ChIP assay was performed in Trim 28 knockdown A549 cells and control (A). ChIP assay was performed in control and Trim28 expressed in A549 cells (B).
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ChIP - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)Image from Ebata KT et al., Epigenetics Chromatin 10(36), Fig 2. Doi: 10.1186/s13072-017-0143-3 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.
ChIP was performed using ab1220. Naïve Mouse ES cells were resuspended in PBS and crosslinked with 1% formaldehyde for 5 minutes at room temperature. Crosslinking was quenched with 125m M glycine. A mouse anti IgG (ab18413) was used as a control. ChIP -seq for H3K9me2 in ES cells treated with and without vitamin C (image a). H3K9me2 signal genome-wide was plotted comparing untreated and vitamin C-treated cells (image b). Fold change for average H3K9me2 signal at gene promoters. Most gene promoters display a reduction in H3K9me2 in vitamin C-treated ES cells (image c). ChIP-qPCR for H3K9me2 in ES cells with or without vitamin C at gene promoters. ChIP for IgG was performed as a negative control (image d). Asterisks represent P < 0.05 by t test
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Western blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)This image is courtesy of an anonymous abreview.Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220) at 1/1000 dilution + HEK293 at 30 µg
Secondary
HRP conjugated polyclonal Sheep anti-Mouse IgG at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 17 kDaBlocking buffer: 5% milk
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
ab1220 staining human kidney sections by IHC-P using EXPOSE IHC detection kit (ab80436). Formalin fixed paraffin embedded tissue sections were pre-treated using heat mediated antigen retrieval (using a pressure cooker) with sodium citrate buffer (pH6) for 30 mins. The section was incubated with ab1220, 5µg/ml, for 1 hour at room temperature. DAB was used as the chromogen and the section was counterstained with haematoxylin and mounted with DPX.
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ELISA using ab1220 at varying antibody concentrations.
The purple line indicates binding to the Human Histone H3 (di methyl K9) peptide ab1772. Binding to the following peptides was not seen:
Human Histone H3 (unmodified ) peptide (ab2903),
Human Histone H3 (mono methyl K9) peptide (ab1771),
Human Histone H3 (tri methyl K9) peptide (ab1773),
Human Histone H3 (di methyl K27) peptide (ab1781),
Human Histone H3 (tri methyl K27) peptide (ab1782),
Human Histone H3 (mono methyl K4) peptide (ab1340),
Human Histone H3 (di methyl K4) peptide (ab7768),
Human Histone H3 (tri methyl K4) peptide (ab1342).This indicates the specificity of ab1220 for di methyl K9 of Histone H3.
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Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220) at 1 µg/ml + HeLa (Human epitherlial carcinoma cell line) Whole Cell Lysate at 20 µg
Secondary
Goat polyclonal to Mouse IgG-H&L- Pre-Adsorbed (HRP) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 17 kDa
Observed band size: 17 kDa
Exposure time: 5 minutesLane 1 : Marker.
All blocking and antibody incubation steps were done with 5% milk in 20mM Tris-HCL, and 0.1% TWEEN-20.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)This image is a courtesy of Anonymous Abreview
ab1220 staining Histone H3 (di methyl K9) in Mouse Tissue sections (Liver, P5) by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 5% BSA for 1 hour at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/100 in PBS/Triton 0,05%) for 3 hours at 25°C. An Alexa Fluor® 488 conjugated Goat anti-mouse (1/250) was used as the secondary antibody.
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Western blot - Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)This image is courtesy of an anonymous AbreviewAll lanes : Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220) at 2.5 µg/ml
Lane 1 : Fruit fly embryo tissue lysate - nuclear at 4.5 µg
Lane 2 : Fruit fly embryo tissue lysate - nuclear at 1.5 µg
Lane 3 : Fruit fly embryo tissue lysate - nuclear at 0.5 µg
Secondary
All lanes : HRP-conjugated Goat anti-mouse IgG polyclonal at 1/2000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 17 kDa
Observed band size: 15 kDa why is the actual band size different from the predicted?
Exposure time: 1 minute
参考文献 (683)
ab1220 は 683 報の論文で使用されています。
- Nicolau M et al. The plant mobile domain proteins MAIN and MAIL1 interact with the phosphatase PP7L to regulate gene expression and silence transposable elements in Arabidopsis thaliana. PLoS Genet 16:e1008324 (2020). PubMed: 32287271
- Jang JE et al. PERK/NRF2 and autophagy form a resistance mechanism against G9a inhibition in leukemia stem cells. J Exp Clin Cancer Res 39:66 (2020). PubMed: 32293500
- Chen TQ et al. EHMT2/G9a Inhibits Aortic Smooth Muscle Cell Death by Suppressing Autophagy Activation. Int J Biol Sci 16:1252-1263 (2020). PubMed: 32174799
- Shamsi F et al. FGF6 and FGF9 regulate UCP1 expression independent of brown adipogenesis. Nat Commun 11:1421 (2020). PubMed: 32184391
- Jozi M et al. Induced DNA hypomethylation by Folic Acid Deprivation in Bovine Fibroblast Donor Cells Improves Reprogramming of Somatic Cell Nuclear Transfer Embryos. Sci Rep 10:5076 (2020). PubMed: 32193457