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Descripción

The anomalous folding and polymerization of the β-amyloid (Aβ) peptide is thought to initiate the neurodegenerative cascade in Alzheimer's disease pathogenesis1. Aβ is predominantly a 40- or 42-amino acid peptide that is prone to self-aggregation into β-sheet-rich amyloid fibrils that are found in the cores of cerebral senile plaques in Alzheimer's disease. Increasing evidence suggests that low molecular weight, soluble Aβ multimers are more toxic than fibrillar Aβ amyloid2. The identification and quantification of low- and high-molecular weight multimeric Aβ species in brain tissue is an essential objective in Alzheimer's disease research, and the methods employed also can be applied to the identification and characterization of toxic multimers in other proteopathies3. Naturally occurring Aβ multimers can be detected by SDS-polyacrylamide gel electrophoresis followed by immunoblotting with Aβ-specific antibodies. However, the separation and detection of multimeric Aβ requires the use of highly concentrated cortical homogenates and antigen retrieval in small pore-size nitrocellulose membranes. Here we describe a technique for the preparation of clarified human cortical homogenates, separation of proteins by SDS-PAGE, and antigen-epitope retrieval/Western blotting with antibody 6E10 to the N-terminal region of the Aβ peptide. Using this protocol, we consistently detect Aβ monomers, dimers, trimers, tetramers, and higher molecular weight multimers in cortical tissue from humans with Alzheimer's pathology.

Pertenece a

PubMed Central (PMC3 - NLM DTD)  

Autor(es)

Rosen, Rebecca F. -  Tomidokoro, Yasushi -  Ghiso, Jorge A. -  Walker, Lary C. - 

Id.: 55213330

Idioma: inglés  - 

Versión: 1.0

Estado: Final

Palabras claveJoVE Neuroscience - 

Tipo de recurso: Text  - 

Tipo de Interactividad: Expositivo

Nivel de Interactividad: muy bajo

Audiencia: Estudiante  -  Profesor  -  Autor  - 

Estructura: Atomic

Coste: no

Copyright: sí

Requerimientos técnicos:  Browser: Any - 

Fecha de contribución: 24-abr-2012

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Otros recursos del mismo autor(es)

  1. Neurodegenerative Diseases: Expanding the Prion Concept The prion paradigm has emerged as a unifying molecular principle for the pathogenesis of many age-re...
  2. Sequential Amyloid-β Degradation by the Matrix Metalloproteases MMP-2 and MMP-9* Background: Abnormal accumulation of Aβ in the brain is associated with neurodegeneration in Alzheim...
  3. Mitochondrial dysfunction induced by a post-translationally modified amyloid linked to a familial mutation in an alternative model of neurodegeneration Familial British dementia (FBD) is an early-onset non-amyloid-β (Aβ) cerebral amyloidosis that prese...
  4. Development of transgenic rats producing human β-amyloid precursor protein as a model for Alzheimer's disease: Transgene and endogenous APP genes are regulated tissue-specifically

    Abstract

    Background

    Alzheimer's disease (AD) is a devastating neurodegenerative dis...

  5. Aβ seeds resist inactivation by formaldehyde Cerebral β-amyloidosis can be exogenously induced by the intracerebral injection of brain extracts c...

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