使用敲除细胞株进行验证

Anti-macroH2A.1抗体[14G7] (ab91528)

概述

  • 产品名称
    Anti-macroH2A.1抗体[14G7]
    参阅全部 macroH2A.1 一抗
  • 描述
    小鼠单克隆抗体[14G7] to macroH2A.1
  • 经测试应用
    适用于: WB, ELISA, Flow Cytmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    Recombinant full length protein of Human macroH2A.1.

  • 阳性对照
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; HepG2; MCF7
  • 常规说明

    This antibody clone is manufactured by Abcam.

性能

  • 形式
    Liquid
  • 存放说明
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
  • 存储溶液
    Preservative: 0.02% Sodium Azide
    Constituents: PBS, pH 7.4
  • Concentration information loading...
  • 纯度
    IgG fraction
  • 克隆
    单克隆
  • 克隆编号
    14G7
  • 同种型
    IgG1
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab91528 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB Use a concentration of 10 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).
ELISA Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

靶标

  • 功能
    Variant histone H2A which replaces conventional H2A in a subset of nucleosomes where it represses transcription. 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. Involved in stable X chromosome inactivation. Inhibits the binding of transcription factors and interferes with the activity of remodeling SWI/SNF complexes. Inhibits histone acetylation by EP300 and recruits class I HDACs, which induces an hypoacetylated state of chromatin. In addition, isoform 1, but not isoform 2, binds ADP-ribose and O-acetyl-ADP-ribose, and may be involved in ADP-ribose-mediated chromatin modulation.
  • 组织特异性
    Ubiquitous.
  • 序列相似性
    Contains 1 histone H2A domain.
    Contains 1 Macro domain.
  • 翻译后修饰
    Monoubiquitinated at either Lys-116 or Lys-117. May also be polyubiquitinated. Ubiquitination is mediated by the CUL3/SPOP E3 complex and does not promote proteasomal degradation. Instead, it is required for enrichment in inactive X chromosome chromatin.
  • 细胞定位
    Nucleus. Chromosome. Enriched in inactive X chromosome chromatin and in senescence-associated heterochromatin.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Core histone macro h2a.1 antibody
    • Core histone macro-H2A.1 antibody
    • H2A histone family member Y antibody
    • H2A.y antibody
    • H2A/y antibody
    • H2AF12M antibody
    • H2AFJ antibody
    • H2afy antibody
    • H2AY_HUMAN antibody
    • Histone H2A.Y antibody
    • Histone macroH2A1 antibody
    • Histone macroH2A1.1 antibody
    • Histone macroH2A1.2 antibody
    • Macroh2a1 antibody
    • MACROH2A1.1 antibody
    • MacroH2A1.2 antibody
    • Medulloblastoma antigen MU MB 50.205 antibody
    • Medulloblastoma antigen MU-MB-50.205 antibody
    • mH2a antibody
    • mH2A1 antibody
    see all

Anti-macroH2A.1 antibody [14G7] 图像



  • Predicted band size : 40 kDa

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: MacroH2A.1 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: HepG2 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab91528 observed at 40 kDa. Red - loading control, ab176560, observed at 50 kDa.

    ab91528 was shown to recognize MacroH2A.1 in wild type cells as signal was lost at the expected MW in MacroH2A.1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and MacroH2A.1 knockout samples were subjected to SDS-PAGE. ab91528 and ab176560 (Rabbit anti-alpha Tubulin loading control) were incubated overnight at 4°C at 10 µg/ml and 1/10000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Overlay histogram showing HeLa cells stained with ab91528 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab91528, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • All lanes : Anti-macroH2A.1 antibody [14G7] (ab91528) at 10 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 40 kDa
    Observed band size : 40 kDa
    Additional bands at : 14 kDa,18 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 2 minutes

Anti-macroH2A.1 antibody [14G7] (ab91528)参考文献

ab91528 has not yet been referenced specifically in any publications.

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