Product nameAnti-Huntingtin antibody [EP867Y]
See all Huntingtin primary antibodies
DescriptionRabbit monoclonal [EP867Y] to Huntingtin
Tested applicationsSuitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
Unsuitable for: IP
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human Huntingtin aa 550-650. The exact sequence is proprietary. Corresponding to residues specific to the apopain cleavage site.
- ICC/IF: SK-N-SH cells. WB: HAP1, SH-SY5Y and HeLa whole cell lysates. IHC-P: Human brain tissue. Flow Cytometry: SH-SY5Y cells.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
Concentration information loading...
PurityTissue culture supernatant
- Anti-Huntingtin antibody [EP867Y] (Alexa Fluor® 488) (ab206048)
- Anti-Huntingtin antibody [EP867Y] (Alexa Fluor® 647) (ab206049)
- Anti-Huntingtin antibody [EP867Y] (Allophycocyanin) (ab224969)
- Anti-Huntingtin antibody [EP867Y] (Phycoerythrin) (ab224970)
- Anti-Huntingtin antibody [EP867Y] - BSA and Azide free (ab225573)
KO cell lysates
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.
|WB||1/10000. Predicted molecular weight: 348 kDa.|
|IHC-P||Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ICC/IF||1/50 - 1/100.|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
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.
modificationsCleaved 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
- AI256365 antibody
- C430023I11Rik antibody
- HD antibody
All lanes : Anti-Huntingtin antibody [EP867Y] (ab45169) at 1/10000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : HTT knockout HeLa cell lysate
Lane 3 : Wild-type HAP1 cell lysate
Lane 4 : HTT knockout HAP1 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 348 kDa
Observed band size: 348 kDa
Lanes 1-4: Merged signal (red and green). Green - ab45169 observed at 348 kDa. Red - loading control ab7291 observed at 50 kDa.
ab45169 Anti-Huntingtin antibody [EP867Y] was shown to specifically react with Huntingtin in wild-type HeLa cells. Loss of signal was observed when knockout sample ab256946 was used. Wild-type and Huntingtin knockout samples were subjected to SDS-PAGE. ab45169 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 10000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Huntingtin knockout HAP1 whole cell lysate (20 µg)
Lane 3: SH-SY5Y 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 specifically recognize Huntingtin in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when Huntingtin knockout samples were examined. 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 1/10,000 dilution and 1/10,000 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 1/10,000 dilution for 1 hour at room temperature before imaging.
Ab45169 staining human Huntingtin in human brain tissue by immunohistochemistry using paraffin embedded tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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?
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) »