Overview

  • Product nameAnti-Huntingtin antibody [EPR5526]
    See all Huntingtin primary antibodies
  • Description
    Rabbit monoclonal [EPR5526] to Huntingtin
  • Tested applicationsFlow Cyt, IHC, ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Huntingtin aa 1-100.
    Database link: P42858

  • Positive control
    • WB: SH-SY5Y, HeLa, PC-12 and Neuro-2a whole cell lysates and mouse and rat brain lysates. IHC-P: Human cerebral cortex tissue, Human astrocytoma tissue, Mouse testis and Rat testis tissue. ICC/IF: Neuro-2a and SH-SY5Y cell lines. Flow cytometry: SH-SY5Y cell line.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    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.

    Alternative versions available:

    Anti-Huntingtin antibody (Alexa Fluor® 488) [EPR5526] (ab197506)

    Anti-Huntingtin antibody (Alexa Fluor® 647) [EPR5526] (ab196931)

Properties

  • FormLiquid
  • Storage instructionsShipped 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 bufferpH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 59% PBS, 0.05% BSA
  • PurityTissue culture supernatant
  • Clonality Monoclonal
  • Clone numberEPR5526
  • IsotypeIgG
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab109115 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
Flow Cyt 1/250.
IHC Use at an assay dependent concentration. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF 1/1000.
WB 1/5000. Detects a band of approximately 348 kDa (predicted molecular weight: 348 kDa).
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • FunctionMay play a role in microtubule-mediated transport or vesicle function.
  • Tissue specificityExpressed 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 diseaseDefects 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 similaritiesBelongs to the huntingtin family.
    Contains 10 HEAT repeats.
  • DomainThe 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 localizationCytoplasm. 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
    • IT 15 antibody
    • IT15 antibody
    • OTTMUSP00000026909 antibody
    • ZHD antibody
    see all

Anti-Huntingtin antibody [EPR5526] images

  • All lanes : Anti-Huntingtin antibody [EPR5526] (ab109115) at 1/5000 dilution

    Lane 1 : SH-SY5Y (Human neuroblastoma from bone marrow cells) whole cell lysate
    Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 4 : Neuro-2a (Mouse neuroblastoma cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

    Predicted band size : 348 kDa
    Observed band size : 348 kDa


    Exposure time : 1 second

    Blocking and Diluting buffer and concentration:  5% NFDM /TBST

  • All lanes : Anti-Huntingtin antibody [EPR5526] (ab109115) at 1/50000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Rat brain lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

    Predicted band size : 348 kDa
    Observed band size : 348 kDa


    Exposure time : 30 seconds

    Blocking and Diluting buffer and concentration: 5% NFDM /TBST

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized

    SH-SY5Y (Human neuroblastoma from bone marrow cells) cells labeling Huntingtin with ab109115 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear and cytoplasmic staining on SH-SY5Y cell line.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
    The negative controls are as follows:
    1. ab191472 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

     

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized

    Neuro-2a (Mouse neuroblastoma cells) cells labeling Huntingtin with ab109115 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear and cytoplasmic staining on Neuro-2a cell line.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
    The negative controls are as follows:
    1. ab191472 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

     

  • Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling Huntingtin with ab109115 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Counter stained with Hematoxylin. Nuclear staining on neuron of human cerebral cortex was observed.

  • Immunohistochemical analysis of paraffin-embedded Human astrocytoma labeling Huntingtin with ab109115 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Counter stained with Hematoxylin. Nuclear staining on cancer cells of astrocytoma.

  • Immunohistochemical analysis of paraffin-embedded Mouse testis labeling Huntingtin with ab109115 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Counter stained with Hematoxylin. Cytoplasmic staining on spermatogenic cells of mouse testis.

  • Immunohistochemical analysis of paraffin-embedded Rat testis labeling Huntingtin with ab109115 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500. Counter stained with Hematoxylin. Weak cytoplasmic staining on spermatogenic cells of rat testis.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed SH-SY5Y (Human neuroblastoma from bone marrow cells) cells labeling Huntingtin with ab109115 at 1/250 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

References for Anti-Huntingtin antibody [EPR5526] (ab109115)

This product has been referenced in:
  • Schut MH  et al. Selection and characterization of llama single domain antibodies against N-terminal huntingtin. Neurol Sci 36:429-34 (2015). WB . Read more (PubMed: 25294428) »

See 1 Publication for this product

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The immunogen is from N-term from amino acid XXXX of P42858.

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