Anti-Huntingtin antibody [EP867Y] (ab45169)
- Product nameAnti-Huntingtin antibody [EP867Y]See all Huntingtin primary antibodies ...
- DescriptionRabbit monoclonal [EP867Y] to Huntingtin
- Tested applicationsWB, IHC-P, ICC/IF, Flow Cyt more details
- Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide corresponding to residues specific to the apopain cleavage site of human Huntingtin.
- Positive control
- SKNSH cell line (IF/ICC)
- General notesProduced under U.S. Patent No. 5,675,063.
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
- Storage bufferPBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
- PurityTissue culture supernatant
- Clonality Monoclonal
- Clone numberEP867Y
- Research Areas
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||WB: 1/10000. Predicted molecular weight: 348 kDa.|
|IHC-P||IHC-P: Use at an assay dependent dilution.|
|ICC/IF||ICC/IF: 1/50 - 1/100.|
|Flow Cyt||Flow Cyt: 1/100.|
- 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.
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Anti-Huntingtin antibody [EP867Y] images
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?)
Ab45169 staining human Huntingtin in human brain tissue by immunohistochemistry using paraffin embedded tissue.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.
References for Anti-Huntingtin antibody [EP867Y] (ab45169)
This product has been referenced in:
- Clemen CS et al. Strumpellin is a novel valosin-containing protein binding partner linking hereditary spastic paraplegia to protein aggregation diseases. Brain 133:2920-41 (2010). Read more (PubMed: 20833645) »
- Cho KJ et al. Inhibition of apoptosis signal-regulating kinase 1 reduces endoplasmic reticulum stress and nuclear huntingtin fragments in a mouse model of Huntington disease. Neuroscience 163:1128-34 (2009). Read more (PubMed: 19646509) »