Overview

  • Product name
    Anti-Huntingtin antibody [EP867Y] - BSA and Azide free
    See all Huntingtin primary antibodies
  • Description
    Rabbit monoclonal [EP867Y] to Huntingtin - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    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.

  • Positive control
    • ICC/IF: SKNSH cells.
  • General notes

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®). 

    ab225573 is a PBS-buffered version of ab45169, containing no BSA or sodium azide, ideal for antibody labeling. Please refer to ab45169 for information on protocols, dilutions, and image data.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.20
    Constituent: PBS
  • Concentration information loading...
  • Purity
    Affinity purified
  • Clonality
    Monoclonal
  • Clone number
    EP867Y
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab225573 in the following tested applications.

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

Application Abreviews Notes
WB Use at an assay dependent concentration. Predicted molecular weight: 348 kDa.
IHC-P Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IP.
  • Target

    • Function
      May play a role in microtubule-mediated transport or vesicle function.
    • Tissue specificity
      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.
    • Involvement in disease
      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).
    • Sequence similarities
      Belongs to the huntingtin family.
      Contains 10 HEAT repeats.
    • Domain
      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.
    • Post-translational
      modifications
      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.
    • Cellular localization
      Cytoplasm. Nucleus. The mutant Huntingtin protein colocalizes with AKAP8L in the nuclear matrix of Huntington's disease neurons.
    • Information by UniProt
    • Database links
    • Alternative names
      • 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

    Images

    • This WB data was generated using the same anti-Huntingtin antibody clone [EP867Y] in a different buffer formulation (cat# ab45169).

      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.

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45169).

    • Ab45169 staining human Huntingtin in human brain tissue by immunohistochemistry using paraffin embedded tissue.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45169).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45169).

    References

    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

    Customer reviews and Q&As

    There are currently no Customer reviews or Questions for ab225573.
    Please use the links above to contact us or submit feedback about this product.

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

    Sign up