This fast track antibody is not yet fully characterized. It is subject to these terms and conditions

製品の概要

  • 製品名Anti-Histone H2B (phospho S32) antibody
    Histone H2B 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Histone H2B (phospho S32)

    This product is a fast track antibody. It has been affinity purified and shows high titre values against the immunizing peptide by ELISA. Read the terms of use »

  • 種交差性
    交差種: Human
    交差が予測される動物種: Mouse
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H2B, phosphorylated at S32.

    (Peptide available as ab18504.)

  • 特記事項


    Serine 32 of Histone H2B was highlighted as being highly conserved in Cheung et al, indicating that it might be a good candidate phosphorylation site.

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Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • 精製度Immunogen affinity purified
  • 一次抗体 備考Serine 32 of Histone H2B was highlighted as being highly conserved in Cheung et al, indicating that it might be a good candidate phosphorylation site.
  • ポリ/モノポリクローナル
  • アイソタイプIgG
  • 研究分野

アプリケーション

Fast track antibodies constitute a diverse group of products that have been released to accelerate your research, but are not yet fully characterized. They have all been affinity purified and show high titre values against the immunizing peptide (by ELISA). Fast track terms of use

アプリケーション Abreviews 特記事項
ICC/IF 1/200.
ELISA Use at an assay dependent concentration. : This antibody gave a positive result in ELISA against the immunizing peptide (ab18504).

ターゲット情報

  • 関連性Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. Linker histones are involved in the formation of higher order structure in chromatin and the maintenance of overall chromatin compaction. Whilst the core histones are highly conserved across a wide range of organisms, the linker histones are less conserved.
  • 細胞内局在Nuclear
  • 参照データベース
  • 別名
    • H2b.1A antibody
    • HIST1H2BD antibody
    • HIST1H2BH antibody
    • HIST1H2BL antibody
    • HIST1H2BM antibody
    • HIST1H2BN antibody
    • HIST2H2BE antibody
    • Histone H2B antibody
    • Histone H2B type 1-D antibody
    • Histone H2B type 1-H antibody
    • Histone H2B type 1-L antibody
    • Histone H2B type 1-M antibody
    • Histone H2B type 1-N antibody
    • Histone H2B type 2-E antibody
    see all

Anti-Histone H2B (phospho S32) antibody 画像

This Fast-Track antibody is not yet fully characterised. These images represent inconclusive preliminary data.

  • All lanes : Anti-Histone H2B (phospho S32) antibody (ab10476) at 1/500 dilution

    Lane 1 : Hela control
    Lane 2 : Gamma irraditaed hela 1h
    Lane 3 : Gamma irraditaed hela 2h
    Lane 4 : Hela control with Human Histone H2B (phospho S32) peptide (ab18504) at 1 µg/ml
    Lane 5 : Gamma irraditaed hela 1h with Human Histone H2B (phospho S32) peptide (ab18504) at 1 µg/ml
    Lane 6 : Gamma irraditaed hela 2h with Human Histone H2B (phospho S32) peptide (ab18504) at 1 µg/ml
    Lane 7 : Hela control with Human Histone H2B peptide (ab18507) at 1 µg/ml
    Lane 8 : Gamma irraditaed hela 1h with Human Histone H2B peptide (ab18507) at 1 µg/ml
    Lane 9 : Gamma irraditaed hela 2h with Human Histone H2B peptide (ab18507) at 1 µg/ml

    Lysates/proteins at 25 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

    Performed under reducing conditions.

    Observed band size : 15 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute

    Lane 1 : Hela control
    Lane 2 : Gamma irraditaed hela 1h
    Lane 3 : Gamma irraditaed hela 2h
    Lane 4 : Hela control + Histone H2B peptide - phospho S32 (ab18504)
    Lane 5 : Gamma irraditaed hela 1h at + Histone H2B peptide - phospho S32 (ab18504)
    Lane 6 : Gamma irraditaed hela 2h at + Histone H2B peptide - phospho S32 (ab18504)
    Lane 7 : Hela control + Histone H2B peptide (ab18507)
    Lane 8 : Gamma irraditaed hela 1h + Histone H2B peptide (ab18507)
    Lane 9 : Gamma irraditaed hela 2h + Histone H2B peptide (ab18507)

    All lysates at 25 ug
    All peptides at 1 ug/ml


    Primary (Lanes 1-9)
    Rabbit polyclonal to Histone H2B (phospho S32) (ab10476) at 1/500 dilution

    Secondary
    Goat polyclonal to Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

    Performed under reducing conditions.

    Observed band size : 15kD
    Exposure time : 1 minute

    This antibody detects a band at just over 15kDa corresponding to H2B on gamma irradiated and control Hela cell lysate. This band is blocked by the immunising phospho peptide (ab18504) but not by the non-phospho peptide (ab18507). This strongly indicates that the antibody is specific for the phosphorylation. We cannot explain the reduction in signal at 1hr after gamma irradiation and and the presence of other higher molecular weight bands. The latter presumably represent cross-reactivity of the antibody with other proteins.

  • ICC/IF image of ab10476 stained HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab10476, 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). This antibody also gave a positive IF result in HepG2, Hek293 and MCF7 cells fixed in 4% PFA at 1ug/ml and Hela, Hek293, HepG2 and MCF7 cells fixed in 100% methanol at 1ug/ml. However, this Fast-Track antibody is not yet fully characterised. This image represents inconclusive preliminary data.
  • IHC image of ab10476 staining in Human Breast adenocarcinoma 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 ab10476, 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.

    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.
  • ab10476 (1/200) staining Histone H2B phospho S32 in Hela cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus (red). for further experimental details please refer to Abreview.

    See Abreview

Anti-Histone H2B (phospho S32) antibody (ab10476) 使用論文

This product has been referenced in:
  • Lau AT  et al. Phosphorylation of histone H2B serine 32 is linked to cell transformation. J Biol Chem : (2011). WB, ICC/IF ; Human . Read more (PubMed: 21646345) »
  • Tjeertes JV  et al. Screen for DNA-damage-responsive histone modifications identifies H3K9Ac and H3K56Ac in human cells. EMBO J 28:1878-89 (2009). WB ; Human . Read more (PubMed: 19407812) »

See all 2 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton X100
Username

Dr. Kirk McManus

Verified customer

投稿 Sep 08 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"