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Quantitative Gel Electrophoresis

  • Victoria J. Mansour
  • Jens R. CoorssenEmail author
Chapter

Abstract

Gel electrophoresis is the most widely used technique for the analysis of protein samples, and there are a variety of methods that can be used to investigate single proteins as well as highly complex protein mixtures. To ensure reproducible and reliable separations of such samples and the resolution of distinct protein species, there has been a substantial amount of research dedicated to optimising methods to the refined techniques available today. There are thus a number of factors that have a marked influence on the practice of quantitative proteomics. The procedures and reagents involved in preparing a protein sample can have a significant effect on the composition of the proteome and/or its resolution by electrophoresis. Furthermore, since most proteins are colourless, a protein stain is required to detect the resolved proteome; thus, it is essential that the characteristics of the stain enable optimal detection regardless of protein type and/or concentration. Notably, to obtain reliable quantitative data, the approach by which images are acquired is equally important.

Keywords

Gel electrophoresis Protein stain Fluorescence Sample preparation Quantitative analysis Protein detection Deep imaging 1D/2D/3D gel electrophoresis 

Abbreviations

1DE

One-dimensional electrophoresis

2DE

Two-dimensional electrophoresis

cCBB

Colloidal Coomassie brilliant blue

CBB

Coomassie brilliant blue

CHAPS

3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate

DIGE

Difference gel electrophoresis

DNA

Deoxyribonucleic acid

DTT

Dithiothreitol

LLD

Lowest limit of detection

LDR

Linear dynamic range

IEF

Isoelectric focussing

IPV

Inter-protein variability

IRF

Infrared fluorescence

MS

Mass spectrometry

MW

Molecular weight

NCCB

Neuhoff colloidal Coomassie brilliant blue

PAGE

Polyacrylamide gel electrophoresis

RNA

Ribonucleic acid

SDS

Sodium dodecyl sulphate

SR

SYPRO Ruby

TBP

Tributyl phosphine

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Copyright information

© Springer International Publishing AG 2018

Authors and Affiliations

  1. 1.School of Medicine, Western Sydney UniversityPenrithAustralia
  2. 2.Faculty of Applied Health Sciences and Faculty of Mathematics and ScienceBrock UniversitySt CatharinesCanada

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