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Isotopic Tracer Techniques in Preclinical ResearchTracer Technique in Basic Research

Isotopic Tracer Techniques in Preclinical Research: Tracer Technique in Basic Research [Tracer techniques remain instrumental in advancing our current understanding of the structure and function of nucleic acids and proteins and their interactions. In this chapter, we will focus on classic protocols that are relevant and pivotal to current research. Besides the radioactive isotopes, the chapter also outlines recent methods that utilize nonradioactive (stable isotope) tracer techniques in proteomic/metabolomic characterization. The radioactive tracer protocols included in this chapter encompass analysis of (a) DNA (Southern blotting), (b) RNA (Northern blotting), (c) DNA-protein interaction (EMSA), and the popular (d) pulse-chase technique to evaluate protein synthesis. Next, the chapter also illustrates the stable isotope protocols which include two versatile techniques employed in cell culture (SILAC) and ex vivo tissue/body fluid analysis (iTRAQ). Each protocol section of the chapter begins with a brief background, overall description of the method, the principle underlying the technique, reagents/equipment required, and a step-wise protocol. At the end of each protocol/technique, one or more study questions are included to improve the clarity and enhance the grasp of the subject. Collectively, techniques described in this chapter are essential to determine the biochemical and molecular mechanisms of the regulation of nucleic acids and proteins in normal and abnormal cellular physiology.] http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png

Isotopic Tracer Techniques in Preclinical ResearchTracer Technique in Basic Research

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Publisher
Springer International Publishing
Copyright
© Springer Nature Switzerland AG 2022
ISBN
978-3-030-99699-4
Pages
7 –44
DOI
10.1007/978-3-030-99700-7_2
Publisher site
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Abstract

[Tracer techniques remain instrumental in advancing our current understanding of the structure and function of nucleic acids and proteins and their interactions. In this chapter, we will focus on classic protocols that are relevant and pivotal to current research. Besides the radioactive isotopes, the chapter also outlines recent methods that utilize nonradioactive (stable isotope) tracer techniques in proteomic/metabolomic characterization. The radioactive tracer protocols included in this chapter encompass analysis of (a) DNA (Southern blotting), (b) RNA (Northern blotting), (c) DNA-protein interaction (EMSA), and the popular (d) pulse-chase technique to evaluate protein synthesis. Next, the chapter also illustrates the stable isotope protocols which include two versatile techniques employed in cell culture (SILAC) and ex vivo tissue/body fluid analysis (iTRAQ). Each protocol section of the chapter begins with a brief background, overall description of the method, the principle underlying the technique, reagents/equipment required, and a step-wise protocol. At the end of each protocol/technique, one or more study questions are included to improve the clarity and enhance the grasp of the subject. Collectively, techniques described in this chapter are essential to determine the biochemical and molecular mechanisms of the regulation of nucleic acids and proteins in normal and abnormal cellular physiology.]

Published: May 4, 2022

Keywords: Southern blotting; Northern blotting; Electrophoretic Mobility Shift Assay (EMSA); Pulse-chase; Stable Isotope Labeling by Amino acids in Cell culture (SILAC); Isobaric Tag for Relative and Absolute Quantitation (iTRAQ)

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