Bioinformatics is a scientific discipline that is still being defined. To some, it is the development of new computer programs that use statistics to discern the relationships between DNA and protein sequences. To others, it is the development and implementation of databases to store and pro ...

The location of an enzyme’s active site is usually known prior to the determination of its three-dimensional structure. Analysis of sequence homology may indicate residues that are near an enzyme’s active site. Catalytic residues are often identified when site-directed mutagenesis e ...

Embryonic stem (ES) cells have a pluripotent ability to differentiate into a variety of cell lineages of all three embryonic germ layers in vitro. The hanging drop culture of ES cell suspension in the absence of leukemia inhibitory factor induces aggregation and differentiation of the cells i ...

Photoaffinity labeling is the process of covalently crosslinking a photoactive ligand, which can be detected postcoupling to a receptor or binding protein. This technique has proven useful in the identification of novel proteins or to further study and characterize known proteins. P ...

Microsatellite instability has been shown to be relevant to various human diseases, including fragile X syndrome (1) and Huntington’s disease (2). In several human cancers, it has been reported that an increase or decrease in the number of repeat units between lymphocyte and tumor DNA derived f ...

Angiogenesis, the formation of new blood vessels from preexisting vessels, is an essential component of many normal biological processes such as embryonic development, wound healing, and endometrial maturation in premenopausal women (1–3). This process is similar to, but not identi ...

The identification of protein binding partners often facilitates understanding of protein complex function. However, identifying binding partners has proven difficult because proteins are often bound to insoluble structures or are only present during certain stages of the ce ...

The in vitro growth of keratinocytes has proved to be an important tool in the study of the normal biology and disease processes involving the skin, e.g., the influence of extrinsic regulators of growth and differentiation, effects of pharmacological agents, dermo-epidermal interactio ...

As delineated in Chapter 26, the directed evolution of enantioselective enzymes for use in synthetic organic chemistry requires the availability of high-throughput screening systems for determining the enantiomeric excess (ee) of thousands of samples (1). In this chapter the prot ...

Adipose tissues (ATs) in mammals are distinguished as being brown or white. Brown adipose tissues (BAT) has as its function the production of heat, and thus has a high oxidative capacity, evidenced by the extraordinarily high density of mitochondria in the cells. White adipose tissue (WAT) is pri ...

Embryonic stem (ES) cells, the undifferentiated cells of early embryos are established as permanent lines (1,2) and are characterized by their self-renewal capacity and the ability to retain their developmental capacity in vivo (3) and in vitro (4–6). The pluripotent properties of ES cells a ...

In vitro cell culture is a necessary prerequisite in acquiring a thorough understanding of the biology and behavior of the cells of interest and is a critical first step in developing cellular therapies. Somatic stem cell biology is concerned with stem cells of the adult and how they may be utilized in ...

cAMP is a ubiquitous second messenger that controls numerous cellular events including movement, growth, metabolism, contraction, and synaptic plasticity. With the emerging concept of compartmentalization of cAMP-dependent signaling, a detailed study of the spatio-tempo ...

The difference between peptides and proteins (the subject of Chapter 2) is that peptides are molecules too small to have a “globular” structure. This means that the spectral assignment process is often much simpler for peptides than it is for proteins, because there are fewer signals present in pe ...

It was first shown by Newmeyer and colleagues in the 1990s that the molecular events of apoptosis could be reconstituted in vitro using Xenopus egg extracts. When the egg extract is allowed to incubate at room temperature for an extended time, the biochemical events of apoptosis are activated spo ...

The goal of this chapter is to give the reader a concise and easy-to-follow guide to proven transient transfection techniques for primary strains and continuous lines of keratinocytes. The emphasis is on readily available and inexpensive resources that also allow for repeatability and a ...

Lymphocytes arise during ontogeny via a series of increasingly restricted intermediates. Initially, the mesoderm gives rise to hemangioblasts, which can differentiate into endothelial precursors, or hematopoietic stem cells (HSCs). HSCs can either self-renew or different ...

The study of metalloproteins by resonance Raman (RR) spectroscopy began two decades ago, with the publication by Long and coworkers of the first RR spectrum of the iron-sulfur protein rubredoxin (1,2). This simple spectrum, which contained only four bands attributed to the Fe-S stretching and ...

Murine embryonic stem (mES) cells are pluripotent cells derived from the inner cell mass of the preimplantation blastocyst. These cells are primitive and undifferentiated and have the potential to become a wide variety of specialized cell types. Mouse ES cells can be regarded as a versatile b ...

StemBase is a database of gene expression data obtained from stem cells and derivatives mainly from mouse and human using DNA microarrays and Serial Analysis of Gene Expression. Here, we describe this database and indicate ways to use it for the study the expression of particular genes in stem cells ...