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RabMAb

Anti-Huntingtin抗体[EP867Y] (ab45169)

概述

  • 产品名称
    Anti-Huntingtin抗体[EP867Y]
    参阅全部 Huntingtin 一抗
  • 描述
    兔单克隆抗体[EP867Y] to Huntingtin
  • 经测试应用
    适用于: WB, IHC-P, ICC/IF, Flow Cytmore details
    不适用于: IP
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Huntingtin aa 550-650. The exact sequence is proprietary. Corresponding to residues specific to the apopain cleavage site.

  • 阳性对照
    • ICC/IF: SKNSH cells.
  • 常规说明

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    This product is a recombinant rabbit monoclonal antibody.

性能

  • 形式
    Liquid
  • 存放说明
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • 存储溶液
    PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
  • 纯度
    Tissue culture supernatant
  • 克隆
    单克隆
  • 克隆编号
    EP867Y
  • 同种型
    IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab45169 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 1/10000. Predicted molecular weight: 348 kDa.
IHC-P Use at an assay dependent concentration.
ICC/IF 1/50 - 1/100.
Flow Cyt 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

  • 应用说明
    Is unsuitable for IP.
  • 靶标

    • 功能
      May play a role in microtubule-mediated transport or vesicle function.
    • 组织特异性
      Expressed in the brain cortex (at protein level). Widely expressed with the highest level of expression in the brain (nerve fibers, varicosities, and nerve endings). In the brain, the regions where it can be mainly found are the cerebellar cortex, the neocortex, the striatum, and the hippocampal formation.
    • 疾病相关
      Defects in HTT are the cause of Huntington disease (HD) [MIM:143100]. HD is an autosomal dominant neurodegenerative disorder characterized by involuntary movements (chorea), general motor impairment, psychiatric disorders and dementia. Onset of the disease occurs usually in the third or fourth decade of life and symptoms progressively worsen leading to death in 10 to 20 years. Onset and clinical course depend on the degree of poly-Gln repeat expansion, longer expansions resulting in earlier onset and more severe clinical manifestations. HD affects 1 in 10,000 individuals of European origin. Neuropathology of Huntington disease displays a distinctive pattern with loss of neurons, especially in the caudate and putamen (striatum).
    • 序列相似性
      Belongs to the huntingtin family.
      Contains 10 HEAT repeats.
    • 结构域
      The N-terminal Gln-rich and Pro-rich domain has great conformational flexibility and is likely to exist in a fluctuating equilibrium of alpha-helical, random coil, and extended conformations.
    • 翻译后修饰
      Cleaved by apopain downstream of the polyglutamine stretch. The resulting N-terminal fragment is cytotoxic and provokes apoptosis.
      Forms with expanded polyglutamine expansion are specifically ubiquitinated by SYVN1, which promotes their proteasomal degradation.
    • 细胞定位
      Cytoplasm. Nucleus. The mutant Huntingtin protein colocalizes with AKAP8L in the nuclear matrix of Huntington's disease neurons.
    • Information by UniProt
    • 数据库链接
    • 别名
      • AI256365 antibody
      • C430023I11Rik antibody
      • HD antibody
      • HD protein antibody
      • HD_HUMAN antibody
      • HDH antibody
      • HTT antibody
      • Huntingtin antibody
      • HUNTINGTON CHOREA antibody
      • Huntington disease protein antibody
      • Huntington's disease protein homolog antibody
      • IT 15 antibody
      • IT15 antibody
      • OTTMUSP00000026909 antibody
      • ZHD antibody
      see all

    Anti-Huntingtin antibody [EP867Y] 图像



    • Predicted band size : 348 kDa

      Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: Huntingtin knockout HAP1 whole cell lysate (20 µg)
      Lane 3: SH-SY-5Y whole cell lysate (20 µg)
      Lane 4: HeLa whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab45169 observed at 348 kDa. Red - loading control, ab18058, observed at 130 kDa.

      ab45169 was shown to recognize Huntingtin when Huntingtin knockout samples were used, along with additional cross-reactive bands. Wild-type and Huntingtin knockout samples were subjected to SDS-PAGE. ab45169 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 10000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 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.

    • Ab45169 staining human Huntingtin in human brain tissue by immunohistochemistry using paraffin embedded tissue.
    • Overlay histogram showing SH-SY5Y cells stained with ab45169 (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 (ab45169, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) ( 1µg/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.

    • ICC/IF image of ab45169 stained SKNSH cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab45169, 1/200 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Anti-Huntingtin antibody [EP867Y] (ab45169) at 1/10000 dilution + SH-SY-5Y cell lysate

      Predicted band size : 348 kDa
      Observed band size : 300 kDa (why is the actual band size different from the predicted?)

    Anti-Huntingtin antibody [EP867Y] (ab45169)参考文献

    This product has been referenced in:
    • Lin CL  et al. Oral treatment with herbal formula B307 alleviates cardiac failure in aging R6/2 mice with Huntington's disease via suppressing oxidative stress, inflammation, and apoptosis. Clin Interv Aging 10:1173-87 (2015). IHC-P ; Mouse . Read more (PubMed: 26229452) »
    • Czeredys M  et al. Expression of genes encoding the calcium signalosome in cellular and transgenic models of Huntington's disease. Front Mol Neurosci 6:42 (2013). WB ; Human . Read more (PubMed: 24324398) »

    See all 4 Publications for this product

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