Anti-Histone H2A 抗体 - ChIP Grade (ab18255)
Key features and details
- Rabbit polyclonal to Histone H2A - ChIP Grade
- Suitable for: ICC/IF, WB, IP, ChIP
- Reacts with: Mouse, Cow, 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 -
アプリケーション
適用あり: ICC/IF, WB, IP, ChIPmore details -
種交差性
交差種: Mouse, Cow, Human -
免疫原
Synthetic peptide within Human Histone H2A aa 100 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
(Peptide available asab19751) -
ポジティブ・コントロール
- WB: HeLa nuclear extract. Calf thymus histone preparation. Histone H2A Recombinant Protein. ChIP: Chromatin from U-2 OS cells. IP: Histone H2A IP in HeLa whole cell lysate. ICC/IF: HeLa cells.
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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
Constituents: 98.98% PBS, 1% BSA
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...
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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ab18255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 |
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ICC/IF | (4) |
1/200. see Abreview submitted by Kirk McManus
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WB | (13) |
Use a concentration of 1 µg/ml. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa).
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IP |
Use a concentration of 5 µg/ml.
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ChIP | (2) |
Use at an assay dependent concentration. Every new batch of this antibody is tested at Abcam in ChIP
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特記事項 |
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ICC/IF
1/200. see Abreview submitted by Kirk McManus |
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa). |
IP
Use a concentration of 5 µg/ml. |
ChIP
Use at an assay dependent concentration. Every new batch of this antibody is tested at Abcam in ChIP |
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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ChIP - Anti-Histone H2A antibody - ChIP Grade (ab18255)Reamon-Buettner and Borlak PLoS One. 2012;7(6):e38531. doi: 10.1371/journal.pone.0038531. Epub 2012 Jun 6. Fig 4. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Myc affects the incorporation levels of histone variant H2A.Z.
qChIP was performed using specific antibodies recognizing A. H2A.Z, B. H2A.Zac, Panel D. H2A, E. H2AK5ac and G. H1. All the qChIP values are expressed as % of input and normalized for total histone H3, with the exception of C and F, where H2A.Z acetylation is noramlized for H2A.Z density, and H2AK5 acetylation is normalized for H2A density, respectively. The box plots show the fold change distribution of each acetylated residue for the two subpopulations.
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All lanes : Anti-Histone H2A antibody - ChIP Grade (ab18255) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg
Lane 3 : Histone H2A Recombinant Protein at 0.1 µg
Lane 4 : Histone H3.1 Recombinant Protein at 0.1 µg
Lane 5 : Histone H4 Recombinant Protein at 0.1 µg
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 14 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutes -
Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A antibody - ChIP Grade (ab18255)Colpitts et al PLoS One. 2011;6(9):e24365. doi: 10.1371/journal.pone.0024365. Epub 2011 Sep 1. Fig 2.
DENV C colocalizes with histones in Huh7 liver cells.
DEN2 C colocalized with H2A (A), H2B (B), H3 (C) and H4 (D) in Huh7 cells. Cells were transfected with GFP-DEN2 C and fixed in 4% paraformaldehyde 48 h post-transfection. Cells were stained with antibodies against histones and a TRITC secondary antibody. Cells were counterstained with DAPI to visualize the nucleus. GFP-DEN2 C expression is green, histone staining is red and DAPI is blue.
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ICC/IF image of ab18255 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.
The cells were fixed in 100% methanol (5 min) then permeabilized using 0.1% PBS-Triton and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to further permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab18255 at 1 µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43 µM for 1 hour at room temperature.
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Histone H2A - ChIP Grade was immunoprecipitated using 0.5 mg HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell extract, 5 µg of Rabbit polyclonal to and 50 µL of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10 min, HeLa whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10 min under agitation.
Proteins were eluted by addition of 40 µL SDS loading buffer and incubated for 10 min at 70°C; 10 µL of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab18255.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 14kDa, non specific bands - 42kDa: We are unsure as to the identity of this extra band; Histone H2A - ChIP Grade
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Chromatin was prepared from U-2 OS (Human bone osteosarcoma epithelial cell line) 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, 6 µL of ab18255 (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 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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Western blot - Anti-Histone H2A antibody - ChIP Grade (ab18255)Colpitts et al PLoS One. 2011;6(9):e24365. doi: 10.1371/journal.pone.0024365. Epub 2011 Sep 1. Fig 5. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
DENV expression disrupts histone oligomerization.
Panel A and B shown:
Huh7 cells were transfected with DEN2 C and/or infected with DEN2 24 h post-transfection. Cells were lysed 24 h post-infection (48 h post-transfection) and lysates were run on 4–12% SDS-PAGE gel. Gels were used in a Western blotting assay with antibodies against histones H2A ab18255 (A), H2B (B), H3 (C) and H4 (D); monomers, dimers and octamers are indicated. Gels were stripped and reprobed with an antibody against actin as a protein loading control. The same amount of protein was loaded in each lane for each gel as a control for expression.
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All lanes : Anti-Histone H2A antibody - ChIP Grade (ab18255) at 1 µg/ml
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) lysate at 20 µg
Lane 2 : HeLa nuclear lysate at 20 µg
Lane 3 : Calf thymus histone lysate at 20 µg
Lane 4 : HeLa lysate at 1 µg/ml with Human Histone H2A peptide (ab19751) at 1 µg/ml
Lane 5 : HeLa nuclear lysate at 1 µg/ml with Human Histone H2A peptide (ab19751) at 1 µg/ml
Lane 6 : Calf thymus histone lysate at 1 µg/ml with Human Histone H2A peptide (ab19751) at 1 µg/ml
Predicted band size: 14 kDa
Observed band size: 14 kDa
Additional bands at: 22 kDa (possible cross reactivity)ab18255 is partially blocked by the immunizing peptide ab19751. There is an additional band at 22kDa in HeLa lysate which is attributed to cross-reactivity.
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Western blot - Anti-Histone H2A antibody - ChIP Grade (ab18255)This image is courtesy of an Abreview submitted by Ragnhild EskelandAll lanes : Anti-Histone H2A antibody - ChIP Grade (ab18255) at 1/1000 dilution
Lane 1 : Native recombinant octamers K562 cells
Lane 2 : Recombinant Human octamers containing H2A
Lane 3 : Recombinant Human octamers containing H2A.Z.2.1
Lane 4 : Recombinant Human octamers containing H2A.Z.1
Lysates/proteins at 0.5 µg per lane.
Secondary
All lanes : HRP-conjugated donkey anti-rabbit IgG polyclonal at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 14 kDa
Observed band size: 15 kDa why is the actual band size different from the predicted?
Exposure time: 5 minutes
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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SDS download
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Datasheet download
参考文献 (130)
ab18255 は 130 報の論文で使用されています。
- Li Q et al. Differential requirement for BRCA1-BARD1 E3 ubiquitin ligase activity in DNA damage repair and meiosis in the Caenorhabditis elegans germ line. PLoS Genet 19:e1010457 (2023). PubMed: 36716349
- Buzova D et al. Detection of cell-free histones in the cerebrospinal fluid of pediatric central nervous system malignancies by imaging flow cytometry. Front Mol Biosci 10:1254699 (2023). PubMed: 38028540
- Liu K et al. Multiomics analysis of naturally efficacious lipid nanoparticle coronas reveals high-density lipoprotein is necessary for their function. Nat Commun 14:4007 (2023). PubMed: 37414857
- Melters DP et al. Single molecule analysis of CENP-A chromatin by high-speed atomic force microscopy. Elife 12:N/A (2023). PubMed: 37728600
- Sapp N et al. HIV-1 Preintegration Complex Preferentially Integrates the Viral DNA into Nucleosomes Containing Trimethylated Histone 3-Lysine 36 Modification and Flanking Linker DNA. J Virol 96:e0101122 (2022). PubMed: 36094316