Protein Arrays: Methods and Protocols is an introduction to protein array technology and its application to the multiplexed detection of proteins. Although protein array technology has some roots in gene array technology, it can only be described as a distant relative. Unlike DNA, with its established rules of base pairing, and therefore predictable biochemical behavior, proteins are rich with diversity. Proteins can be large or small, compact or extended, basic or acidic, hydrophobic or hydrophilic, and so on. Just as importantly, their behavior is determined by the environment in which they reside, and so the composition of the buffer in which experiments are performed has a dramatic impact on the outcome of the experiment. Thus, if the goal is to simultaneously measure the expression of a large number of proteins, these variables must be addressed. Not to be deterred, scientists have created a variety of solutions to successfully detect and ch- acterize multiple proteins simultaneously. It is the intent of this volume to introduce to the reader a set of technological solutions to the diversity problem as well as to provide the reader with some examples of practical applications of these technologies.
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Protein arrays make possible the detection and quantitation of many proteins simultaneously, thus enabling researchers to ask fundamental questions about biological processes and to discover biomarkers that can be used diagnostically. In Protein Arrays: Methods and Protocols, innovative experimentalists describe in detail the methods they have developed to synthesize and construct protein arrays for basic and clinical research. The authors present protocols to create and immobilize the capture substrate-the first task in designing a protein array-using a variety of affinity capture reagents, including antibodies, peptides, aptamers, biotin, chemical reagents, and chromatographic substrates. Once synthesized, these arrays can be used to analyze protein-protein interactions and posttranslational modifications, such as phosphorylation, as well as to discover and characterize potential diagnostic markers. Protocols to accomplish these tasks are also presented. The protocols presented follow the successful Methods in Molecular Biology™ series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Diverse and highly practical, Protein Arrays: Methods and Protocols offers basic and clinical investigators a broad spectrum of approaches to the generation of protein arrays, as well as their uses in biomarkers discovery, in assay development, in clinical sample testing, in signal transduction analysis and characterization, and for creating the next generation of molecular tools.
"Diverse and highly practical...offers basic and clinical investigators a broad spectrum of approaches to the generation of protein arrays..." - Clinical Laboratory International
"...provides a broad spectrum of approaches for the generation, production and application of protein arrays." - BTi
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