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Protocol
Protein Phosphatase Type 1 and Type 2A Assays
Protein phosphatases type 1 (PP1) and type 2A (PP2A) are the only activities known in mammalian tissues to dephosphorylate glycogen phosphorylase a. Phosphorylase was the first enzyme demonstrated to undergo regu...
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Protocol
Expression of Heterologous Integrin Genes
Cell-extracellular matrix interactions are mediated predominantly by integrins, cell-surface receptors that exist as heterodimers of noncovalently associated α and β subunits (1). Integrins have been implicated i...
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Protocol
Electrophysiological Studies on Receptors In Vitro
In the past 20 years that in vitro preparations of central nervous system (CNS) cells have been used to access excitable membrane properties with electrophysiological techniques, there have been remarkable cha...
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Protocol
Functional Analysis of Cytoskeletal Components in the Develo** Visual System of Drosophila
A central issue in neuroscience is to understand the mechanisms underlining the stereotyped patterns of neuronal connectivity. Studies of neuronal development in both vertebrates and invertebrates have establi...
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Protocol
Intraocular Delivery of Recombinant Virus
In the past 5 yr, advances in technology have been made that allow efficient somatic transfer of functional genes to target cells in the eye in vivo. The ability to deliver functional genes to these cells is d...
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Protocol
Transfection Assays for Transformation of Model Colonic Cell Lines
The recognition of cancer as a genetic disease has changed the approach investigators take to understanding the mechanisms of carcinogenesis. The discovery of oncogenes, and the recognition of the inactivation...
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Protocol
Labeling Stem Cells In Vitro for Identification of Their Differentiated Phenotypes After Grafting into the CNS
Stem cells have unlimited therapeutic potential for restoring CNS function lost secondary to trauma or degenerative disease. To date, stem cell grafts have only in a few instances partially ameliorated functio...
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Protocol
Derivation and Propagation of Embryonic Stem Cells in Serum- and Feeder-Free Culture
The availability of murine embryonic stem (ES) cells has revolutionized the study of mammalian development and disease. We recently developed a culture medium that has enabled us to identify the essential sign...
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Protocol
μParaflo™ Biochip for Nucleic Acid and Protein Analysis
We describe in this chapter the use of oligonucleotide or peptide microarrays (arrays) based on microfluidic chips. Specifically, three major applications are presented: (1) microRNA/small RNA detection using ...
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Protocol
Construction of Infectious Clones of Double-Stranded DNA Viruses of Plants Using Citrus Yellow Mosaic Virus as an Example
Double-stranded DNA (dsDNA) viruses of plants are believed to be plant pararetroviruses. Their genome is replicated by reverse transcription of a larger than unit-length terminally redundant RNA transcript of ...
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Protocol
Labeling Stem Cells In Vitro for Identification of their Differentiated Phenotypes after Grafting into the CNS
Grafting neural stem cells is a widely used experimental approach to central nervous system (CNS) repair after trauma or neurodegeneration. It is likely to be a realistic clinical therapy for human CNS disorde...
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Protocol
Analysis of Microdissected Cells by Two-Dimensional LC-MS Approaches
Laser capture microdissection (LCM) is a powerful tool that enables the isolation of specific cell types from tissue sections, overcoming the problem of tissue heterogeneity and contamination. We combined the ...
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Protocol
Isolation and Primary Culture of Human Proximal Tubule Cells
Primary cultures of renal proximal tubule cells (PTC) have been widely used to investigate tubule cell function. They provide a model system where confounding influences of renal haemodynamics, cell heterogene...
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Protocol
Qualitative and Quantitative Analysis of Peptide Microarray Binding Experiments Using SVM-PEPARRAY
A main objective of analyzing peptide array-based binding experiments is to uncover the relationship between a peptide sequence and the binding outcome. Limited by the peptide array technologies available for ...
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Protocol
Measurement of Glomerular Filtration Rate in Conscious Mice
Glomerular filtration rate (GFR) is an important index of renal function and routinely used in patient care and basic research to evaluate progression of renal diseases or test the efficacy of novel therapeuti...
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Protocol
Web-Based Design of Peptide Microarrays Using μPepArray Pro
Peptide microarrays (peptide arrays) have increasingly become an important research tool for studying protein detection, profiling, and protein–protein interactions, and they have the potential to foster high-...
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Protocol
Bacterial Delivery of siRNAs: A New Approach to Solid Tumor Therapy
RNAi is a powerful research tool for specific gene silencing and may also lead to promising novel therapeutic strategies. However, the development of RNAi-based therapies has been slow due to the lack of targe...
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Protocol
Purification of Arabidopsis Argonaute Complexes and Associated Small RNAs
Argonaute (AGO) proteins recruit small RNAs to form effector complexes of RNA interference (RNAi), collectively termed RNA-induced silencing complexes (RISCs). Here, we describe detailed protocols for the puri...
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Protocol
Covalent Conjugation of Multi-walled Carbon Nanotubes with Proteins
Linkage of proteins to carbon nanotubes (CNTs) is fundamentally important for applications of CNTs in medicinal and biological fields, as well as in biosensor or chemically modulated nanoelectronic devices. In...
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Protocol
Characterization of Anti-competitor Activities Produced by Oral Bacteria
Most bacteria in nature exist in multispecies communities known as biofilms. In the natural habitat where resources (nutrient, space, etc.) are usually limited, individual species must compete or collaborate w...
