Authors:
Is a unique compendium that offers a complete solution of protocols on GAPDH protein analysis for researchers at all levels including undergraduates, graduates,postgraduates and research scientists in life science /general and applied biology
Outlines exceptional methods to analyze enzymic proteins (e.g. GAPDH) under native, non-denatured condition by immunoblotting
Illustrates both qualitative and quantitative analysis of GAPDH as an enzyme and as a non-enzymic protein
Encompasses methods for the analysis of recently discovered isoform, secretory-GAPDH
Offers notes/tips that are essential for the successful completion of protocols
Provides nice illustrations to support the text
Buy it now
Buying options
Tax calculation will be finalised at checkout
Other ways to access
This is a preview of subscription content, log in via an institution to check for access.
Table of contents (10 chapters)
-
Front Matter
-
Back Matter
About this book
This book presents modern and classic analytical approaches that are crucial for the biochemical and functional characterization of the archetypal protein, glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The distinguishing feature of the book is that it covers, in addition to other methods, some of the uncommon but valuable techniques as well. For example, in-gel visualization of enzyme activity, immunoblotting protocols for native (non-denatured) proteins, and proteins resolved by pH-gradient [IEF-isoelectrofocusing], etc. These expedient methods are relevant and vital for the verification of biochemical properties of GAPDH, or similar protein of interest.
This work outlines detailed protocols that are essential to investigate classical (cellular) and recently reported extracellular (secretory) isoforms of GAPDH. Precisely, the book covers techniques pertinent to enzymatic and non-enzymatic analysis of GAPDH that include, but not limited to, electrophoretic mobility shift assay (EMSA), two-dimensional (2D)-immunoblotting, immunofluorescence/confocal microscopy, mass spectrometry, ion-exchange and affinity chromatography. Â
Readers will discover the importance of the experimental methods described in the book as they relate to the evaluation of the role and significance of GAPDH. Furthermore, majority of the methods described in the book have also been validated in the author’s laboratory, besides other research groups worldwide, underlining the repeatability and reproducibility of the protocols. Each method begins with an abstract and a brief background emphasizing its application and relevance. This will enable the readers to determine the choice of experimental design according to their research objectives.
The book explains the methods systematically with ample illustrations to facilitate quick and easy comprehension of the practical knowledge. Although the book is focused on GAPDH, many of the protocols may be adopted to other proteins or enzymes with minimal modifications. Noteworthy, it is unequivocally established that GAPDH is a multifunctional protein involved in several cellular processes of health & disease conditions. Hence, this book will be a valuable practical guide for young researchers, scientists and clinician-scientists.
Authors and Affiliations
-
Dept. of Radiology & Radiological Sciences, Johns Hopkins University School of Medicine, Baltimore, USA
Shanmugasundaram Ganapathy-Kanniappan
Bibliographic Information
Book Title: Advances in GAPDH Protein Analysis: A Functional and Biochemical Approach
Authors: Shanmugasundaram Ganapathy-Kanniappan
DOI: https://doi.org/10.1007/978-981-10-7342-7
Publisher: Springer Singapore
eBook Packages: Biomedical and Life Sciences, Biomedical and Life Sciences (R0)
Copyright Information: Springer Nature Singapore Pte Ltd. 2017
Hardcover ISBN: 978-981-10-7340-3Published: 29 January 2018
Softcover ISBN: 978-981-13-5621-6Published: 11 December 2018
eBook ISBN: 978-981-10-7342-7Published: 16 January 2018
Edition Number: 1
Number of Pages: XIV, 115
Number of Illustrations: 23 b/w illustrations, 12 illustrations in colour
Topics: Protein Science