Protocols

Primary Astroglial Cultures From Mouse Cerebral Cortex

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Submitted by: Josep Saura
Posted Date: 07 Sep 2010

Brief Description

This protocol describes how to prepare primary cultures consisting of >95% astrocytes from neonatal mouse cerebral cortex. It also works for late embryos, for rat and for most CNS regions. Protocols for preparing mixed glial or highly enriched astroglial cultures are also available in this database.

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Protocols

Magnetofection Transfection of Murine Embryonic Motor Neurons

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Submitted by: Wilfried Rossoll
Posted Date: 06 Jun 2010

Brief Description

A protocol for the efficient transfection of cultured primary motor neurons via magnetofection, a transfection technology based on the delivery of DNA-coated magnetic nanobeads. This protocol uses the MF10000 Super Magnetic Plate or MF14000 Mega Magnetic Plate (Oz Biosciences) and NM51000 NeuroMag transfection reagent (Oz Biosciences).

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Protocols

Alzheimer's Association Flow Chart for Lumbar Puncture CSF Processing

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Submitted by: Kaj Blennow
Posted Date: 18 Nov 2009

Brief Description

The objective of the QC program is to standardize CSF biomarker measurements between labs. This protocol is a standardized protocol for lumbar puncture and CSF sample processing.Notes:For the CSF processing and analysis part, it is recommended that the clinical laboratories in this program adhere to national quality guidelines and have received a national (or international) accreditation for medical laboratories.

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Protocols

Biofluid Sample Collection Protocol for ADNI

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Submitted by: John Trojanowski
Posted Date: 21 Oct 2009

Brief Description

This protocol includes procedures for the collection, processing and shipment of clinical laboratory and APOE samples, cell immortalization samples at baseline, blood and urine samples for biomarkers, and cerebral spinal fluid samples. For more information and updated information see the ADNI website.

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Protocols

Plasma Fluid Collection and Processing

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Submitted by: David Holtzman
Posted Date: 28 Sep 2009

Brief Description

An overview of how to collect human plasma and process it for future analysis.

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Protocols

Cerebrospinal Fluid (CSF) Collection and Processing

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Submitted by: David Holtzman
Posted Date: 28 Sep 2009

Brief Description

An overview of how to collect CSF by lumbar puncture and process it for future analysis.

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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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