Anti-Histone H2A 抗体 - ChIP Grade (ab116677)

製品の概要

  • 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
  • アプリケーション適用あり: ChIP, IHC-P, ICC/IF, IP, WBmore details
  • 種交差性
    交差種: Human
  • 免疫原

    Synthetic peptide conjugated to KLH, corresponding to a region within C terminal amino acids 101-130 of Human Histone H2A (NP_003507.1, NP_001035807.1).

  • ポジティブ・コントロール
    • K562 cell line lysate IHC-P: Human normal colon FFPE tissue sections. ICC/IF: Hela cells

製品の特性

関連製品

アプリケーション

Our Abpromise guarantee covers the use of ab116677 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
ChIP Use 5 µg for 25 µg of chromatin.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 2 µg/ml.
IP Use a concentration of 5 µg/ml.
WB 1/100 - 1/500. Predicted molecular weight: 14 kDa.

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 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
  • 参照データベース
  • 別名
    • H2a 615 antibody
    • H2A antibody
    • H2A GL101 antibody
    • H2A histone family member A antibody
    • H2A.1 antibody
    • H2A.2 antibody
    • H2A/a antibody
    • H2A/m antibody
    • H2A/O antibody
    • H2A/q antibody
    • H2A1B_HUMAN antibody
    • H2AFA antibody
    • H2AFE antibody
    • H2AFL antibody
    • H2AFM antibody
    • H2AFO antibody
    • H2AFQ antibody
    • HIST1H2AE antibody
    • HIST1H2AJ antibody
    • HIST2H2AA antibody
    • HIST2H2AA3 antibody
    • HIST2H2AB antibody
    • HIST2H2AC antibody
    • Histone 1 H2ae antibody
    • Histone 2 H2aa3 antibody
    • Histone 2 H2ab antibody
    • Histone 2 H2ac antibody
    • Histone H2A type 1 B antibody
    • Histone H2A type 1 C antibody
    • Histone H2A type 1 E antibody
    • Histone H2A type 1 J antibody
    • Histone H2A type 1-B/E antibody
    • Histone H2A.2 antibody
    • Histone H2A/a antibody
    • Histone H2A/m antibody
    • MGC74460 antibody
    see all

Anti-Histone H2A antibody - ChIP Grade 画像

  • Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab116677 (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.

  • Histone H2A was immunoprecipitated using 0.5mg HeLa whole cell extract, 5µg of Rabbit polyclonal to and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, HeLa whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab116677.

    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).

    Band: 14kDa; Histone H2A

  • IHC image of ab116677 staining Histone H2A in human colon formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. 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 ab116677, 5µg/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. No primary antibody was used in the negative control (shown on the inset).

     

    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.

  • ab116677 staining Histone H2A in HeLa cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab116677 at 2μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 2 μg/ml (shown in green). AlexaFluor®350 WGA was used at a 1/200 dilution and incubated for 1h with the cells, to label plasma membranes (shown in blue). Nuclear DNA was labelled in red with 1.25 μM DRAQ5™ (ab108410), which was added to the secondary antibody mixture. A secondary only negative control is displayed, which indicates that the Histone H2A staining observed is due to primary antibody specificity and not to unspecific binding of the secondary antibody to the cells.

  • Anti-Histone H2A antibody - ChIP Grade (ab116677) at 1/100 dilution + K562 cell line lysate at 35 µg

    Predicted band size : 14 kDa

Anti-Histone H2A antibody - ChIP Grade (ab116677) 使用論文

ab116677 has not yet been referenced specifically in any publications.

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