| Protocol: |
96 Well Abeta ELISA Using Fluorescent Substrate |
| Method: |
ELISA |
| Description: |
96 Well Abeta ELISA Using Fluorescent Substrate |
| Lab: |
The Laboratory of Dennis J. Selkoe |
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| Protocol: |
Abeta ELISA |
| Method: |
ELISA |
| Description: |
A protocol to detect Abeta in tissue culture supernatant and in homogenized tissue. |
| Lab: |
The Laboratory of Dennis J. Selkoe |
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| Protocol: |
ACE Activity Assay |
| Method: |
Protein-Enzyme Assay |
| Description: |
A protocol to measure angiotensin-converting enzyme (ACE) activity. |
| Lab: |
The Laboratory of Dennis J. Selkoe |
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| Protocol: |
Acid Phosphatase Histochemistry for Mouse Brain |
| Method: |
TBD |
| 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. |
| Lab: |
Nixon |
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| Protocol: |
Alzheimer's Association Flow Chart for Lumbar Puncture CSF Processing |
| Method: |
Collection of Biofluids , Protein-Enzyme Assay |
| 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. |
| Lab: |
Clinical Neurochemistry Laboratory - Göteborg University, Sahlgren's University Hospital, Mölndal, Sweden |
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| Protocol: |
Biofluid Sample Collection Protocol for ADNI |
| Method: |
Collection of Biofluids |
| Description: |
This section includes procedures for the collection, processing and shipment of clinical laboratory and APOE samples at screen, cell immortalization samples at baseline, blood and urine samples for biomarkers, and lastly cerebral spinal fluid samples. |
| Lab: |
Laboratory of John Trojanowski |
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| Protocol: |
Cerebrospinal Fluid Collection and Processing |
| Method: |
Collection of Biofluids |
| Description: |
Cerebrospinal Fluid Collection and Processing |
| Lab: |
Laboratory of Anne Fagan |
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| Protocol: |
Coating of Transwell Filters |
| Method: |
Cell Culture, TBD |
| Description: |
Protocol for coating of Transwell Filters. |
| Lab: |
The Laboratory of Dennis J. Selkoe |
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| Protocol: |
Collagenase/Dispase Mouse Brain Isolation |
| Method: |
Cell Culture |
| Description: |
Collagenase/Dispase Mouse Brain Isolation. |
| Lab: |
The Laboratory of Dennis J. Selkoe |
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| Protocol: |
Coupling Antibodies to Protein A/G |
| Method: |
Protein Isolation/Purification |
| Description: |
This protocol can be used to generate an affinity matrix consisting of an antibody covalently linked to Protein A/G-coated beads. |
| Lab: |
The Laboratory of Dennis J. Selkoe |
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| Protocol: |
Culturing Embryonic Fibroblasts |
| Method: |
Cell Culture |
| Description: |
Culturing Embryonic Fibroblasts. |
| Lab: |
The Laboratory of Dennis J. Selkoe |
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| Protocol: |
DNA Ethanol Precipitation |
| Method: |
Nucleic Acid Isolation/Purification |
| Description: |
DNA ethanol precipitation protocol. |
| Lab: |
The Laboratory of Dennis J. Selkoe |
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| Protocol: |
Efficient Transfection of Murine Embryonic Motor Neurons via Magnetofection |
| Method: |
Cell Culture, Gene Expression , Transfection |
| Description: |
An optimized protocol for the efficient transfection of cultured primary motor neurons via magnetofection, a novel transfection technology based on the delivery of DNA-coated magnetic nanobeads. |
| Lab: |
Bassell Lab |
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| Protocol: |
Extraction of Abeta From Brain Tissues for Application to Abeta ELISA |
| Method: |
Protein Isolation/Purification |
| Description: |
This protocol is a method to extract Abeta from brain tissues for application to Abeta ELISA. |
| Lab: |
Laboratory for Proteolytic Neuroscience, RIKEN Brain Science Institute |
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| Protocol: |
Extraction of Human Abeta from APP Tg mouse brains |
| Method: |
Protein Isolation/Extraction |
| Description: |
Extraction of human Abeta from APP Tg mouse brains into TBS-soluble, guanidine HCI-soluble fractions |
| Lab: |
The Laboratory of Dennis J. Selkoe |
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| Protocol: |
Horseradish Peroxidase (HRP) Uptake/Trafficking |
| Method: |
TBD |
| 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. |
| Lab: |
Nixon |
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| Protocol: |
IHC (Immunoperoxidase) |
| Method: |
Immunohistochemistry |
| Description: |
IHC (immunoperoxidase) protocol. |
| Lab: |
The Laboratory of Dennis J. Selkoe |
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| Protocol: |
Immunocytochemistry and Immunofluorescence for Brain Sections |
| Method: |
Immunohistochemistry , TBD |
| 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. |
| Lab: |
Nixon |
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| Protocol: |
Immunoelectron Microscopy of Brain Tissues Embedded in L R White Resin |
| Method: |
Microscopy |
| Description: |
A protocol on tissue processing and post-embedding immunogold labeling |
| Lab: |
Mayo Clinic Neuropathology Laboratory |
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| Protocol: |
Immunoprecipitation |
| Method: |
Protein Isolation/Purification |
| Description: |
Immunoprecipitation protocol. |
| Lab: |
The Laboratory of Dennis J. Selkoe |
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| Protocol: |
Immunostaining for Neprilysin |
| Method: |
Immunohistochemistry |
| Description: |
This is a protocol for the immunostaining of neprilysin. |
| Lab: |
Laboratory for Proteolytic Neuroscience, RIKEN Brain Science Institute |
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| Protocol: |
In vivo Cathepsin D and Cathepsin B Activity Assay |
| Method: |
TBD |
| Description: |
In vivo cathepsin activity assay can provide information about the lysosomal enzyme activity in live cell condition. |
| Lab: |
Nixon |
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| Protocol: |
MDCK Transwell Filter Monolayers |
| Method: |
Cell Culture |
| Description: |
MDCK Transwell Filter Monolayers. |
| Lab: |
The Laboratory of Dennis J. Selkoe |
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| Protocol: |
Mouse Genotyping Protocols |
| Method: |
Molecular Biology |
| Description: |
Mouse Genotyping Protocols |
| Lab: |
Jackson Laboratory |
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| Protocol: |
Neprilysin Activity Assay |
| Method: |
Protein-Enzyme Assay |
| Description: |
A protocol to detect Abeta in tissue culture. |
| Lab: |
The Laboratory of Dennis J. Selkoe |
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| Protocol: |
Plasma Fluid Collection and Processing |
| Method: |
Collection of Biofluids |
| Description: |
Plasma Fluid Collection and Processing |
| Lab: |
Laboratory of Anne Fagan |
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| Protocol: |
Preparation and Characterization of Toxic Abeta Aggregates for Structural and Functional Studies in Alzheimer's Disease Research |
| Method: |
Protein Preparation |
| Description: |
We describe methods and detailed protocols for reproducibly preparing Aβ aggregates of defined size distribution and morphology, including monomers, protofibrils and fibrils, using size exclusion chromatography. In addition, we describe detailed biophysical procedures for elucidating the structural features, aggregation kinetics and toxic properties of the different Aβ aggregation states, with special emphasis on protofibrillar intermediates. |
| Lab: |
Lashuel Laboratory of Molecular Neurobiology and Neuroproteomics |
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| Protocol: |
Preparation of Mouse Brain for Biochemical and Histological Analysis |
| Method: |
Tissue Preparation |
| Description: |
A protocol for preparation of mouse brain for biochemical and histological analysis. |
| Lab: |
The Laboratory of Dennis J. Selkoe |
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| Protocol: |
Primary Astroglial Cultures From Mouse Cerebral Cortex |
| Method: |
Cell Culture |
| Description: |
This is our protocol 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. Our protocols for preparing mixed glial or highly enriched astroglial cultures are also available at the Alzforum database. |
| Lab: |
Neuroinflammation Lab |
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| Protocol: |
Primary Microglial Cultures From Mouse Cerebral Cortex |
| Method: |
Cell Culture |
| Description: |
This is our protocol to prepare primary cultures consisting of >95% microglia from neonatal mouse cerebral cortex. It also works for late embryos, for rat and for most CNS regions. Our protocols for preparing mixed glial or highly enriched astroglial cultures are also available at the Alzforum database. |
| Lab: |
Neuroinflammation Lab |
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| Protocol: |
Primary Mixed Glial Cultures From Mouse Cerebral Cortex |
| Method: |
Cell Culture |
| Description: |
This is our protocol to prepare primary glial cultures consisting of 75% astrocytes and 25% microglia approximately. We use neonatal mouse cerebral cortex, but the protocol also works for late embryos, for rat, and for most CNS regions. Our protocols for preparing highly enriched astroglial or microglial cultures are also available at the Alzforum database. |
| Lab: |
Neuroinflammation Lab |
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| Protocol: |
Primary Support Cultures of Hippocampal and Substantia Nigra Neurons |
| Method: |
Cell Culture, Tissue Preparation |
| Description: |
Here we present a novel method for culturing 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. |
| Lab: |
Alzheimer's and Parkinson's Disease Laboratory, Brain and Mind Research Institute, University of Sydney |
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| Protocol: |
Pronuclear Injection for the Production of Transgenic Mice |
| Method: |
Transgenic Model |
| 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. Transgenic expression of a truncated form
of the microtubule-associated protein tau (Dtau) is used as an example for the anticipated results.
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| Lab: |
Alzheimer's and Parkinson's Disease Laboratory, Brain and Mind Research Institute, University of Sydney |
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| Protocol: |
Strip & Reprobe Protocol (PVDF membranes) |
| Method: |
Western Blot |
| Description: |
Strip & Reprobe protocol. |
| Lab: |
The Laboratory of Dennis J. Selkoe |
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| Protocol: |
Tau Immunocytochemistry |
| Method: |
Immunohistochemistry |
| Description: |
A widely distributed protocol for tau staining in transgenic mice. |
| Lab: |
Laboratory of Peter Davies at Albert Einstein College of Medicine |
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| Protocol: |
The Alzheimer’s Association multi-center study
on lumbar puncture feasibility
|
| Method: |
Collection of Biofluids , Protein-Enzyme Assay |
| Description: |
A study to gauge the feasibility of routine lumbar puncture for CSF analysis |
| Lab: |
Clinical Neurochemistry Laboratory - Göteborg University, Sahlgren's University Hospital, Mölndal, Sweden |
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| Protocol: |
Thioflavin S Staining |
| Method: |
Histology |
| Description: |
Thioflavin-S staining protocol. |
| Lab: |
The Laboratory of Dennis J. Selkoe |
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| Protocol: |
Western Blotting |
| Method: |
ELISA |
| Description: |
This protocol uses the Bio-Rad blotting system. |
| Lab: |
The Laboratory of Dennis J. Selkoe |
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