Anti-Histone H4 (mono methyl K20) 抗体 - ChIP Grade (ab9051)
Key features and details
- Rabbit polyclonal to Histone H4 (mono methyl K20) - ChIP Grade
- Suitable for: IHC-P, IHC-Fr, Flow Cyt (Intra), WB, ICC/IF, ChIP, ChIP-sequencing, IHC - Wholemount
- Reacts with: Mouse, Cow, Human, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe
- Isotype: IgG
リコンビナント抗体で、ロット間での高い再現性を実現
- 異なるロット間での安定した再現性
- 容易なスケールアップ
- 評価試験による特異性の確認済み
- 倫理基準に準拠 - アニマル・フリーの生産
製品の概要
-
製品名
Anti-Histone H4 (mono methyl K20) antibody - ChIP Grade
Histone H4 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to Histone H4 (mono methyl K20) - ChIP Grade -
由来種
Rabbit -
アプリケーション
適用あり: IHC-P, IHC-Fr, Flow Cyt (Intra), WB, ICC/IF, ChIP, ChIP-sequencing, IHC - Wholemountmore details -
種交差性
交差種: Mouse, Cow, Human, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe
交差が予測される動物種: Mammals -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
特記事項
Interphase: The cells within the culture show a considerable variability in the intensity of staining with the antibody. The relationship between trimethylation levels and cell cycle have not yet been determined but may be a contributor to the amount of methylation detected in each cell. Heterochromatic regions of the interphase nucleus are the primary sites of trimethylation observed by indirect immunofluroescence. Mitosis: Discrete chromosomal regions are labelled intensely, with lower level fluorescence throughout the remainder of the chromosome arms Data from IF performed by Kirk McManus in the lab of Michael Hendzel as part of the antibody characterisation at cellnucleus.com
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As
製品の特性
-
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
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 -
一次抗体 備考
Interphase: The cells within the culture show a considerable variability in the intensity of staining with the antibody. The relationship between trimethylation levels and cell cycle have not yet been determined but may be a contributor to the amount of methylation detected in each cell. Heterochromatic regions of the interphase nucleus are the primary sites of trimethylation observed by indirect immunofluroescence. Mitosis: Discrete chromosomal regions are labelled intensely, with lower level fluorescence throughout the remainder of the chromosome arms Data from IF performed by Kirk McManus in the lab of Michael Hendzel as part of the antibody characterisation at cellnucleus.com -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
-
ChIP Related Products
-
Compatible Secondaries
-
Control Peptide
-
Isotype control
-
Recombinant Protein
-
Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab9051の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
---|---|---|
IHC-P | (2) |
Use a concentration of 0.05 - 0.2 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
|
IHC-Fr | (1) |
Use at an assay dependent concentration.
|
Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody. |
|
WB | (7) |
1/1000.
|
ICC/IF | (5) |
Use at an assay dependent concentration. Cautionary note: At low dilutions, the antibody may cross-react with microtubules. This can be minimized by increasing the dilution of the antibody when used in immunofluroescence.
|
ChIP | (2) |
Use 2-25 µg for 25 µg of chromatin.
|
ChIP-sequencing |
Use at an assay dependent concentration. PubMed: 20200201
|
|
IHC - Wholemount | (1) |
Use at an assay dependent concentration.
|
特記事項 |
---|
IHC-P
Use a concentration of 0.05 - 0.2 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
IHC-Fr
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000. |
ICC/IF
Use at an assay dependent concentration. Cautionary note: At low dilutions, the antibody may cross-react with microtubules. This can be minimized by increasing the dilution of the antibody when used in immunofluroescence. |
ChIP
Use 2-25 µg for 25 µg of chromatin. |
ChIP-sequencing
Use at an assay dependent concentration. PubMed: 20200201 |
IHC - Wholemount
Use at an assay dependent concentration. |
ターゲット情報
-
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 H4 family. -
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.
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. -
細胞内局在
Nucleus. Chromosome. - Information by UniProt
-
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
see all -
別名
- dJ160A22.1 antibody
- dJ160A22.2 antibody
- dJ221C16.1 antibody
see all
画像
-
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 ab9051 (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). Primers and probes are located in the first kb of the transcribed region.
-
All lanes : Anti-Histone H4 (mono methyl K20) antibody - ChIP Grade (ab9051) at 1 µg/ml
Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate (ab121)
Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H4 peptide (ab14963) at 0.5 µg/ml
Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (mono methyl K4) peptide (ab1340) at 0.5 µg/ml
Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (di methyl K4) peptide (ab7768) at 0.5 µg/ml
Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) withHuman Histone H3 (tri methyl K4) peptide (ab1342) at 0.5 µg/ml
Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H4 (mono methyl K20) peptide (ab17043) at 0.5 µg/ml
Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Histone H4 peptide (17-24) - di methyl K20 at 0.5 µg/ml
Lane 8 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H4 (tri methyl K20) peptide (ab17567) at 0.5 µg/ml
Lane 9 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (mono methyl K9) peptide (ab1771) at 0.5 µg/ml
Lane 10 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) withHuman Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µg/ml
Lane 11 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) 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 Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Observed band size: 13 kDa why is the actual band size different from the predicted?
Exposure time: 10 seconds -
IHC image of Histone H4 (mono methyl K20) staining in Human normal 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 ab9051, 0.05µg/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.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
-
Rabbit polyclonal to Histone H4 mono methyl K20 (1/250).
HeLa cells cultured on glass coverslips were fixed with 4% paraformaldehyde and then stained with ab9051 (green). Total chromatin was visualized using DAPI staining (red).
This image is part of the antibody characterisation at www.cellnucleus.com -
Rabbit polyclonal to Histone H4 mono methyl K20 at 1/1000 on histone prep (Sigma).
Peptide competition (shown above lanes) at 1ug/ml.
This peptide blocking study shows that ab9051 is specific for Histone H4 (mono methyl K20). The activity of the antibody is specifically blocked by the addition of the H4 mono methyl peptide (ab17043).
-
Rabbit polyclonal to Histone H4 mono methyl K20 (1/250).
HeLa cells cultured on glass coverslips were fixed with 4% paraformaldehyde and then stained with ab9051 (green). Total chromatin was visualized using DAPI staining (red).
Left: merge; center: DAPI; right: Mono K20
This image is part of the antibody characterisation at www.cellnucleus.com -
Rabbit polyclonal to Histone H4 mono methyl K20 (1/250).
Mouse 10T1 cells cultured on glass coverslips were fixed with 4% paraformaldehyde and then stained with ab9051.
Top left: metaphase cell. Mono methyl K20 (green); DAPI (red)
Top right: interphase cell. Mono methyl K20 (green); DAPI (red)
Lower left: interphase cell. Mono methyl K20 (green); DAPI (blue), TBP (red)
Lower right: interphase cell. Mono methyl K20 (green); DAPI (red)
This image is part of the antibody characterisation at www.cellnucleus.com
プロトコール
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.
データシートおよび資料
-
SDS download
-
Datasheet download
参考文献 (129)
ab9051 は 129 報の論文で使用されています。
- Wang F et al. SETD8 cooperates with MZF1 to participate in hyperglycemia-induced endothelial inflammation via elevation of WNT5A levels in diabetic nephropathy. Cell Mol Biol Lett 27:30 (2022). PubMed: 35350980
- Rang FJ et al. Single-cell profiling of transcriptome and histone modifications with EpiDamID. Mol Cell 82:1956-1970.e14 (2022). PubMed: 35366395
- Stolz P et al. TET1 regulates gene expression and repression of endogenous retroviruses independent of DNA demethylation. Nucleic Acids Res 50:8491-8511 (2022). PubMed: 35904814
- Naughton C et al. Human centromere repositioning activates transcription and opens chromatin fibre structure. Nat Commun 13:5609 (2022). PubMed: 36153345
- Zhang T et al. Replication collisions induced by de-repressed S-phase transcription are connected with malignant transformation of adult stem cells. Nat Commun 13:6907 (2022). PubMed: 36376321