Anti-Histone H2A 抗体 - ChIP Grade (ab15653)
Key features and details
- Rabbit polyclonal to Histone H2A - ChIP Grade
- Suitable for: WB, ICC/IF, ChIP
- Reacts with: Mouse, Human
- 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
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製品名
Anti-Histone H2A antibody - ChIP Grade
Histone H2A 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to Histone H2A - ChIP Grade -
由来種
Rabbit -
特異性
ab15653 was generated using an immunogen specific to the C-terminal of H2A2B (SwissProt Q8IUE6). Interestingly mass spec analysis on immunoprecipitated material using ab15653 indicates that the antibody recognises all isotypes of H2A. -
アプリケーション
適用あり: WB, ICC/IF, ChIPmore details -
種交差性
交差種: Mouse, Human -
免疫原
Synthetic peptide corresponding to Human Histone H2A aa 100 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available asab15660) -
ポジティブ･コントロール
- WB: HeLa nuclear extract, NIH3T3 cells; ICC: HeLa
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特記事項
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
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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
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 -
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
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ChIP Related Products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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ab15653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 |
|---|---|---|
| WB |
Use a concentration of 0.5 µg/ml. Detects a band of approximately 18 kDa (predicted molecular weight: 14 kDa).
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| ICC/IF |
Use a concentration of 1 µg/ml.
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| ChIP |
Use 2-25 µg for µg of chromatin.
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| 特記事項 |
|---|
|
WB
Use a concentration of 0.5 µg/ml. Detects a band of approximately 18 kDa (predicted molecular weight: 14 kDa). |
|
ICC/IF
Use a concentration of 1 µg/ml. |
|
ChIP
Use 2-25 µg for µg of chromatin. |
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 H2A family. -
翻訳後修飾
The chromatin-associated form is phosphorylated on Thr-121 during mitosis.
Deiminated on Arg-4 in granulocytes upon calcium entry.
Monoubiquitination of Lys-120 by RING1 and RNF2/RING2 complex gives a specific tag for epigenetic transcriptional repression and participates in X chromosome inactivation of female mammals. It is involved in the initiation of both imprinted and random X inactivation. Ubiquitinated H2A is enriched in inactive X chromosome chromatin. Ubiquitination of H2A functions downstream of methylation of 'Lys-27' of histone H3. Monoubiquitination of Lys-120 by RNF2/RING2 can also be induced by ultraviolet and may be involved in DNA repair. Following DNA double-strand breaks (DSBs), it is ubiquitinated through 'Lys-63' linkage of ubiquitin moieties by the E2 ligase UBE2N and the E3 ligases RNF8 and RNF168, leading to the recruitment of repair proteins to sites of DNA damage. Monoubiquitination and ionizing radiation-induced 'Lys-63'-linked ubiquitination are distinct events.
Phosphorylation on Ser-2 is enhanced during mitosis. Phosphorylation on Ser-2 by RPS6KA5/MSK1 directly represses transcription. Acetylation of H3 inhibits Ser-2 phosphorylation by RPS6KA5/MSK1.
Symmetric dimethylation on Arg-4 by the PRDM1/PRMT5 complex may play a crucial role in the germ-cell lineage. -
細胞内局在
Nucleus. Chromosome. - Information by UniProt
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参照データベース
- Entrez Gene: 3012 Human
- Entrez Gene: 317772 Human
- Entrez Gene: 8335 Human
- Entrez Gene: 8338 Human
- Entrez Gene: 319166 Mouse
- Omim: 142720 Human
- Omim: 602786 Human
- Omim: 602797 Human
see all -
別名
- H2a 615 antibody
- H2A antibody
- H2A GL101 antibody
see all
画像
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Western blot - Anti-Histone H2A antibody - ChIP Grade (ab15653)All lanes : Anti-Histone H2A antibody - ChIP Grade (ab15653) at 1 µg/ml
Lane 1 : CTH cell lysate at 0.5 µg
Lane 2 : HeLa whole cell lysate at 20 µg
Lane 3 : HeLa Nuclear Triton Enriched cell lysate at 20 µg
Lane 4 : NIH/3T3 whole cell lysate at 20 µg
Lane 5 : NIH/3T3 Nuclear Triton Enriched cell lysate at 20 µg
Lane 6 : Histone H3 Recombinant Protein (Negative control) at 0.1 µg
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Predicted band size: 14 kDa
Additional bands at: 17 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 30 secondsBlocking buffer: 3% Milk
Gel type: MES
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Western blot - Anti-Histone H2A antibody - ChIP Grade (ab15653)All lanes : Anti-Histone H2A antibody - ChIP Grade (ab15653) at 0.5 µg/ml
Lane 1 : HeLa nuclear extract
Lane 2 : HeLa nuclear extract with Human Histone H2A peptide (ab15660) at 1 µg/ml
Predicted band size: 14 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?
ab15653 recognises H2A in HeLa nuclear extracts (lane1), which is successfully blocked using the immunizing peptide (lane2). -
ChIP - Anti-Histone H2A antibody - ChIP Grade (ab15653)Chromatin was prepared from Hela 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 ab15653 (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). Primers and probes are located in the first kb of the transcribed region.
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Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A antibody - ChIP Grade (ab15653)ICC/IF image of ab15653 stained human HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab15653, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
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
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Datasheet download
参考文献 (8)
ab15653 は 8 報の論文で使用されています。
- Sun H et al. miR-4429 sensitized cervical cancer cells to irradiation by targeting RAD51. J Cell Physiol 235:185-193 (2020). PubMed: 31190335
- Kim J et al. Replication Stress Shapes a Protective Chromatin Environment across Fragile Genomic Regions. Mol Cell 69:36-47.e7 (2018). PubMed: 29249653
- Kozlowski M et al. MacroH2A histone variants limit chromatin plasticity through two distinct mechanisms. EMBO Rep 19:N/A (2018). PubMed: 30177554
- Jullien D et al. Chromatibody, a novel non-invasive molecular tool to explore and manipulate chromatin in living cells. J Cell Sci 129:2673-83 (2016). WB ; Human . PubMed: 27206857
- Lee YH et al. Regulation of DNA Damage Response by Estrogen Receptor ß-Mediated Inhibition of Breast Cancer Associated Gene 2. Biomedicines 3:182-200 (2015). PubMed: 28536406
- Loza-Muller L et al. Fibrillarin methylates H2A in RNA polymerase I trans-active promoters in Brassica oleracea. Front Plant Sci 6:976 (2015). Mouse . PubMed: 26594224
- Reitsma JM et al. Human cytomegalovirus IE1 protein disrupts interleukin-6 signaling by sequestering STAT3 in the nucleus. J Virol 87:10763-76 (2013). PubMed: 23903834
- Burd CJ et al. Analysis of chromatin dynamics during glucocorticoid receptor activation. Mol Cell Biol 32:1805-17 (2012). ChIP . PubMed: 22451486
