Human HTT caspase cleavage sites generate fragment-specific forms of the protein. Caspase-3/7 has been shown to generate cleavage sites at animo acids 513 and 552. Caspase-2 cleaves at amino acid 552 and caspase-6 at amino acid 586. Neo-specific antibody ab155930 recognizes the 586 cleaved fragment without detecting the full-length form.
Neoepitope antibodies distinguish smaller cleaved fragments or processed forms of proteins versus the intact full-length or precursor by using a designed peptide purification process to maximize immunoreactivity to a specific cleavage site.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/500. Predicted molecular weight: 348 kDa.
1/50 - 1/200.
1/50 - 1/200.
1/20 - 1/100.
May play a role in microtubule-mediated transport or vesicle function.
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).
Belongs to the huntingtin family. Contains 10 HEAT repeats.
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.
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.
Cytoplasm. Nucleus. The mutant Huntingtin protein colocalizes with AKAP8L in the nuclear matrix of Huntington's disease neurons.
Sandwich ELISA was performed with a monoclonal antibody to Huntingtin and ab155930 to determine the antigen concentration of the Huntingtin cleavage products. The curve represents a dose response for neo586 in 293T cells overexpressing the Huntingtin construct.
Immunofluorescent analysis of formalin fixed, permeabilized 293T cells transfected with Huntingtin23Q (left panel) and Huntingtin148Q (right panel)stop constructs ending in amino acid 586 labeling capase cleaved Huntingtin with ab155930 at 1/50(green). Nuclei were stained with Hoechst 33342 (blue).
Western blot - Anti-Huntingtin antibody (ab155930)
All lanes : Anti-Huntingtin antibody (ab155930) at 1/500 dilution
Lane 1 : Lysate containing endogenous Huntingtin without capase activity Lane 2 : Lysate containing endogenous Huntingtin with capase2 activity Lane 3 : Lysate containing endogenous Huntingtin with capase3 activity Lane 4 : Lysate containing endogenous Huntingtin with capase6 activity Lane 5 : Lysate containing endogenous Huntingtin with capase7 activity Lane 6 : Lysate containing overexpressed recombinant Huntingtin fragment (aa1-513) Lane 7 : Lysate containing overexpressed recombinant Huntingtin fragment (aa1-536) Lane 8 : Lysate containing overexpressed recombinant Huntingtin fragment (aa1-552) Lane 9 : Lysate containing overexpressed recombinant Huntingtin fragment (aa1-586)
Lysates/proteins at 20 µg per lane.
Secondary Lanes 1-7 & 9 : Goat anti-rabbit HRP conjugated antibody at 1/30000 dilution Lane 8 : Goat anti-rabbit HRP conjugated antibody at 1/30000 dilution