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Protocol
Mesoderm Induction in Xenopus
Vertebrate mesoderm forms a diversity of tissues, including notochord, somitic muscle, pronephros, mesenchyme, and blood. In the amphibian Xenopus laevis, mesoderm arises in the equatorial region (marginal zone) ...
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Protocol
Anti-Nitrotyrosine Antibodies for Immunohistochemistry
Nitrotyrosine is an important marker for the formation of peroxynitrite and possibly other reactive nitrogen species derived from nitric oxide in vivo (1). Pathological conditions can substantially increase the p...
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Protocol
Quantitation of Angiotensin II Receptors by Competitive Reverse-Transcription Polymerase Chain Reaction
Becker-Andre and Hahlbrock first described competitive RT-PCR (1). In the competitive RT-PCR, a synthetic RNA/DNA control (competitor), generally called internal standard, is coamplified with the target nucleic a...
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Protocol
Bioluminescence Resonance Energy Transfer Assays for Protein-Protein Interactions in Living Cells
We recently developed a bioluminescence resonance energy transfer (BRET) system for assaying protein-protein interactions (1,2), which has been used successfully for studying the interaction of circadian clock pr...
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Protocol
Complementary DNA Libraries
Complementary DNA libraries reflect gene expression at certain times for specific cells, whereas genomic DNA libraries represent all genetic information in somatic cells. The complexity of cellular organizatio...
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Protocol
mRNA/cDNA Library Construction Using RNA—Polymerase Cycling Reaction
Molecular profiling of single-cell gene expression permits the high-definition investigation of intracellular gene activity and physiological status of the cells under certain special conditions, such as patho...
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Protocol
Subtractive Hybridization for the Identification of Differentially Expressed Genes Using Uraci-DNA Glycosylase and Mung-Bean Nuclease
Subtractive hybridization is a technique for detecting differences of gene expression between two populations of RNAs or complementary DNAs (cDNAs). It is based on base pair complementary that nucleic acid seq...
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Protocol
Subtractive Cloning of Differential Genes Using RNA-PCR
The ability to compare two different messenger RNA (mRNA) or complementary DNA (cDNA) libraries has permitted studies into the role of differentially expressed genes involved in the mechanisms of neoplastic tr...
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Protocol
Assessment of the Quality of mRNA Libraries by Agarose Gel Electrophoresis
Gel electrophoresis has been widely used in separating and purifying macromolecules such as proteins and nucleic acids that differ in size, shape, charge, and conformations. The gel basically acts as a tube in...
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Protocol
Future Perspectives
We have made tremendous strides in our understanding and utilization of the new methodologies in molecular biology over the past 50 yr. With our recent access to complete genome sequences, the function of diff...
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Protocol
Localization of Duck Hepatitis B Virus Polymerase Within Cells
Hepadnaviruses are small, DNA-containing viruses that replicate by reverse transcription (1). They have a lipid envelope surrounding an icosahedral protein core particle, whose shell is composed of a single viral...
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Protocol
Site-Specific Peptide Immobilization Strategies for the Rapid Detection of Kinase Activity on Microarrays
The massive throughput offered by array-based technologies can only be realized with the development of equally powerful strategies that offer reproducible consistency. The competence of arrays and efficacy of...
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Protocol
Construction of Recombinant Vaccinia Viruses Using Leporipoxvirus-Catalyzed Recombination and Reactivation of Orthopoxvirus DNA
Poxvirus DNA is not infectious because the initiation of the infective process requires proteins encapsidated along with the virus genome. However, infectious virus can be produced if purified poxvirus DNA is ...
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Protocol
Site-Specific Immobilization of Biotinylated Proteins for Protein Microarray Analysis
The postgenome era has led to a new frontier of proteomics that requires the development of protein microarray, which enables us to unravel the biological function of proteins in a massively parallel fashion. ...
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Protocol
In Vivo Detection of Protein-Protein Interaction in Plant Cells Using BRET
The emerging technique of bioluminescence resonance energy transfer (BRET) allows us to detect protein interactions in live cells and in real time, thus providing a new window into cellular signal transduction...
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Protocol
Autologous Mesenchymal Stem Cells for Post-Ischemic Myocardial Repair
Cell transplantation is a novel therapy for patients with postinfarction ven tricular dysfunction and congestive heart failure. The potential of mesenchy mal stem cells (MSCs) to be used in the injured myocard...
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Protocol
Cellular Therapy With Autologous Skeletal Myoblasts for Ischemic Heart Disease and Heart Failure
Cardiomyocytes are final differentiated cells that lose the ability to regen erate. Autologous cellular transplantation for cardiac repair has recently emerged as a promising new approach for end-stage heart f...
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Protocol
Site-Specific Immobilization of Proteins in a Microarray
The completion of the Human Genome Project has generated enormous opportunities, as well as challenges, to protein scientists. The key issue now is to develop efficient strategies that allow high-throughput st...
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Protocol
Incorporation of Phosphotyrosyl Mimetic 4- (Phosphonodifluoromethyl)phenylalanine (F2Pmp) Into Signal Transduction-Directed Peptides
Phosphotyrosyl (pTyr)-containing sequences in proteins serve important roles in cellular signal transduction. Often, synthetic pTyr-containing peptides based on cognate sequences surrounding these pTyr residue...
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Protocol
Animal Models of Cardiac Fibrosis
A collagen network, composed largely of type I and III fibrillar collagens, is found in the heart’s interstitial space. This network has multiple functions, including the preservation of tissue architecture an...
