Anti-Histone H4 (di methyl K20, tri methyl K20) 抗体 [6F8-D9] (ab78517)
製品の概要
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製品名Anti-Histone H4 (di methyl K20, tri methyl K20) antibody [6F8-D9]
Histone H4 一次抗体 製品一覧 -
製品の詳細Mouse monoclonal [6F8-D9] to Histone H4 (di methyl K20, tri methyl K20)
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由来種Mouse
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アプリケーション適用あり: IHC-P, WB, ELISA, Dot blot, ICC/IF, Flow Cytmore details
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種交差性交差種: Mouse, Human, Caenorhabditis elegans
交差が予測される動物種: Xenopus laevis, Arabidopsis thaliana, Drosophila melanogaster, Mammal
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免疫原
Synthetic peptide within Human Histone H4 (tri methyl K20). The exact sequence is proprietary.
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特記事項
This antibody clone 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.
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 -
Concentration information loading... -
精製度IgG fraction
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ポリ/モノモノクローナル
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クローン名6F8-D9
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ミエローマx63-Ag8.653
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アイソタイプIgG1
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軽鎖の種類kappa
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研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry kits
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Isotype control
アプリケーション
Our Abpromise guarantee covers the use of ab78517 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 |
|---|---|---|
| IHC-P | Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. | |
| WB | Use at an assay dependent concentration. Predicted molecular weight: 11 kDa. | |
| ELISA | Use at an assay dependent concentration. | |
| Dot blot | Use at an assay dependent concentration. | |
| ICC/IF | Use a concentration of 5 µg/ml. | |
| Flow Cyt | Use 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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 H4 family.
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翻訳後修飾Acetylation at Lys-6 (H4K5ac), Lys-9 (H4K8ac), Lys-13 (H4K12ac) and Lys-17 (H4K16ac) occurs in coding regions of the genome but not in heterochromatin.
Citrullination at Arg-4 (H4R3ci) by PADI4 impairs methylation.
Monomethylation and asymmetric dimethylation at Arg-4 (H4R3me1 and H4R3me2a, respectively) by PRMT1 favors acetylation at Lys-9 (H4K8ac) and Lys-13 (H4K12ac). Demethylation is performed by JMJD6. Symmetric dimethylation on Arg-4 (H4R3me2s) by the PRDM1/PRMT5 complex may play a crucial role in the germ-cell lineage.
Monomethylated, dimethylated or trimethylated at Lys-21 (H4K20me1, H4K20me2, H4K20me3). Monomethylation is performed by SET8. Trimethylation is performed by SUV420H1 and SUV420H2 and induces gene silencing.
Phosphorylated by PAK2 at Ser-48 (H4S47ph). This phosphorylation increases the association of H3.3-H4 with the histone chaperone HIRA, thus promoting nucleosome assembly of H3.3-H4 and inhibiting nucleosome assembly of H3.1-H4.
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. Monoubiquitinated at Lys-92 of histone H4 (H4K91ub1) in response to DNA damage. The exact role of H4K91ub1 in DNA damage response is still unclear but it may function as a licensing signal for additional histone H4 post-translational modifications such as H4 Lys-21 methylation (H4K20me).
Sumoylated, which is associated with transcriptional repression.
Crotonylation (Kcr) is specifically present in male germ cells and marks testis-specific genes in post-meiotic cells, including X-linked genes that escape sex chromosome inactivation in haploid cells. Crotonylation marks active promoters and enhancers and confers resistance to transcriptional repressors. It is also associated with post-meiotically activated genes on autosomes. -
細胞内局在Nucleus. Chromosome.
- Information by UniProt
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参照データベース
- Entrez Gene: 318846 Drosophila melanogaster
- Entrez Gene: 3771854 Drosophila melanogaster
- Entrez Gene: 3771893 Drosophila melanogaster
- Entrez Gene: 3771908 Drosophila melanogaster
- Entrez Gene: 3771935 Drosophila melanogaster
- Entrez Gene: 3771938 Drosophila melanogaster
- Entrez Gene: 3771941 Drosophila melanogaster
- Entrez Gene: 3771947 Drosophila melanogaster
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別名
- H4_HUMAN antibody
- H4F2 antibody
- H4FN antibody
see all
画像
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![Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 (tri methyl K20) antibody [6F8-D9] - ChIP Grade (ab78517)](http://a.abcam.com/ps/products/78/ab78517/Images/ab78517-171911-ab7851734231copy.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 (tri methyl K20) antibody [6F8-D9] - ChIP Grade (ab78517)Image courtesy of Maki Asami, University of Bath, U.K.ICC/IF image of ab78517 stained mouse oocyte. Oocytes were fixed in formaldehyde and then incubated in 3% serum with 0.25% Triton for 30 minutes at 37°C. Cells were incubated with the antibody (ab78517) at 1/200 for 2 hours at 37°C. The secondary antibody used was a polyclonal Goat-anti-mouse conjugated to Alexa Fluor® 488 (1/250).
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![Western blot - Anti-Histone H4 (di methyl K20, tri methyl K20) antibody [6F8-D9] (ab78517)](http://a.abcam.com/ps/products/78/ab78517/Images/ab78517-94324-Dsimageab78517AP21835371.jpg)
Western blot - Anti-Histone H4 (di methyl K20, tri methyl K20) antibody [6F8-D9] (ab78517)All lanes : Anti-Histone H4 (di methyl K20, tri methyl K20) antibody [6F8-D9] (ab78517) at 1 µg/ml
Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate
Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H4 peptide (ab14963) at 0.5 µg/ml
Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate withHuman Histone H3 (mono methyl K4) peptide (ab1340) at 0.5 µg/ml
Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate withHuman Histone H3 (di methyl K4) peptide (ab7768) at 0.5 µg/ml
Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate withHuman Histone H3 (tri methyl K4) peptide (ab1342) at 0.5 µg/ml
Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate withHuman Histone H4 (mono methyl K20) peptide (ab17043) at 0.5 µg/ml
Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H4 peptide (17-24) - di methyl K20 at 0.5 µg/ml
Lane 8 : Calf Thymus Histone Preparation Nuclear Lysate withHuman Histone H4 (tri methyl K20) peptide (ab17567) at 0.5 µg/ml
Lane 9 : Calf Thymus Histone Preparation Nuclear Lysate withHuman Histone H3 (mono methyl K9) peptide (ab1771) at 0.5 µg/ml
Lane 10 : Calf Thymus Histone Preparation Nuclear Lysate withHuman Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µg/ml
Lane 11 : Calf Thymus Histone Preparation Nuclear Lysate withHuman Histone H3 (tri methyl K9) peptide (ab1773) at 0.5 µg/ml
Lysates/proteins at 0.5 µg per lane.
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 11 kDa
Observed band size: 13 kDa (why is the actual band size different from the predicted?)
Exposure time: 90 seconds
ab78517 is specific for Histone H4 (tri-methyl K20). This is illustrated in lane 8 where the activity of ab78517 is specifically blocked by the addition of the immunizing peptide (ab17567). -
![Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 (di methyl K20, tri methyl K20) antibody [6F8-D9] (ab78517)](http://a.abcam.com/ps/products/78/ab78517/Images/Histone-H4-Primary-antibodies-ab78517-1.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 (di methyl K20, tri methyl K20) antibody [6F8-D9] (ab78517)ICC/IF image of ab78517 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 (ab78517, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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 100% methanol fixed (5 min) Hek293, HepG2 and MCF7 cells at 5µg/ml, and in 4% PFA fixed (10 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (di methyl K20, tri methyl K20) antibody [6F8-D9] (ab78517)](http://a.abcam.com/ps/products/78/ab78517/Images/Histone-H4-Primary-antibodies-ab78517-11.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (di methyl K20, tri methyl K20) antibody [6F8-D9] (ab78517)IHC image of Histone H4 (tri methyl K20) staining in normal human skin formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab78517, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. -
![Flow Cytometry-Anti-Histone H4 (tri methyl K20) antibody [6F8-D9] - ChIP Grade(ab78517)](http://a.abcam.com/ps/products/78/ab78517/Images/Histone-H4-Primary-antibodies-ab78517-14.jpg)
Flow Cytometry-Anti-Histone H4 (tri methyl K20) antibody [6F8-D9] - ChIP Grade(ab78517)Overlay histogram showing HeLa cells stained with ab78517 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab78517, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed.
参考文献
This product has been referenced in:
- Flora P et al. Transient transcriptional silencing alters the cell cycle to promote germline stem cell differentiation in Drosophila. Dev Biol 434:84-95 (2018). WB . Read more (PubMed: 29198563) »
- Tikhmyanova N et al. Small molecule perturbation of the CAND1-Cullin1-ubiquitin cycle stabilizes p53 and triggers Epstein-Barr virus reactivation. PLoS Pathog 13:e1006517 (2017). Read more (PubMed: 28715492) »
レビューと Q&A
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