Protocols

Immunohistochemistry for Brain Sections (ABC and Immunofluorescence)

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Submitted by: Ralph Nixon
Posted Date: 19 Jun 2009

Brief Description

Frequently used on human and mouse brain sections for detecting pathologies in the endocytic-autophagic-lysosomal pathways such as changes in the numbers, types and distribution of vacuoles, and expression levels of individual molecules in the pathways.

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Protocols

In vivo Cathepsin D and Cathepsin B Activity Assay

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Submitted by: Ralph Nixon
Posted Date: 19 Jun 2009

Brief Description

In vivo cathepsin activity assay can provide information about the lysosomal enzyme activity in living cells.

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Protocols

Horseradish Peroxidase (HRP) Uptake/Trafficking

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Submitted by: Ralph Nixon
Posted Date: 19 Jun 2009

Brief Description

Uptake of HRP into neuronal cells occurs via bulk endocytosis, especially at axonal terminals. Endocytosed HRP is transported in a retrograde direction and delivered into autophagic-lysosomal vacuoles in the cell body and, therefore, can serve as a marker for studying endocytic trafficking to autophagic-lysosomal compartments.

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Protocols

Acid Phosphatase Histochemistry for Mouse Brain

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Submitted by: Ralph Nixon
Posted Date: 19 Jun 2009

Brief Description

Autophagic-lysosomal compartments contain numerous acid hydrolases. Analysis using enzyme histochemistry for acid phosphatase at the ultrastructural level can provide information about the trafficking and distribution of acid hydrolases in neuronal processes and perikarya and within specific subcellular compartments. We usually use a lead capture technique with cytidine 5’-monophosphate (CMP) as the substrate.

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Protocols

Pronuclear Injection for the Production of Transgenic Mice

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Submitted by: Jürgen Götz
Posted Date: 13 Apr 2009

Brief Description

Pronuclear injection is the most widely used protocol for the generation of transgenic mice. Here, we describe all steps involved from DNA purification to the set up of a mouse colony including vasectomy, injection of the DNA into a donor zygote, transfer of injected zygotes into recipient foster mice, screening of offspring and establishment of transgenic mouse lines. We discuss the use of neuron-specific promoters to express proteins with a role in Alzheimer disease.

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Protocols

Primary Support Cultures of Hippocampal and Substantia Nigra Neurons

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Submitted by: Jürgen Götz
Posted Date: 13 Apr 2009

Brief Description

This protocol describes how to culture embryonic (E16.5) murine hippocampal neurons using a spatially separated ring of cortical neurons for neurotrophic support. This method allows long-term cultures at a very low cell density, and therefore, the study of single embryo preparations and isolated neurons. This method has been adopted for neurons from the substantia nigra (E16.5) with support from a ring of striatal neurons. This protocol is available through Nature Protocols.

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Protocols

96 Well Aβ ELISA Using Fluorescent Substrate

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Submitted by: Dennis Selkoe
Posted Date: 25 Feb 2009

Brief Description

A method for detecting Aβ in human samples

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Protocols

Immunostaining for Neprilysin

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Submitted by: Nobuhisa Iwata
Posted Date: 24 Feb 2009

Brief Description

This is a protocol for the immunostaining of neprilysin using a high temperature antigen unmasking technique and the immunoperoxidase-indirect or fluorescence-indirect tyramide signal amplification (TSA) method.

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Protocols

Extraction of Aβ From Brain for Application to Aβ ELISA

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Submitted by: Nobuhisa Iwata
Posted Date: 24 Feb 2009

Brief Description

This protocol is a method to extract Aβ from brain tissues for application to Aβ ELISA.

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