Two-dimensional sodium dodecyl sulfate

One of the most useful techniques for visualization of the proteome is two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (2-D SDS-PAGE). This technique possesses unmatched resolving power for separation of proteins [2-4] and has been used extensively to analyze proteins [5-8], their regulation [9-18], and posttranslational modifications [19-22], Several tech-... 

Fig. 3. Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of [ S]methionine polypeptides from cholic acid-induced (A) and control (B) cultures of Eubacterium V.P.I. 12708. Arrows indicate the synthesis of new polypeptides.

Separation of the target protein(s) using one- or two-dimensional sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) or HPLC techniques. 

Ramnath, M., Beukes, M., Tamura, K., and Hastings, J.W. (2000). Absence of a putative mannose-specific phosphotransferase system enzyme IIAB component in a leucocin A-resistant strain of Listeria monocytogenes, as shown by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Appl Environ Microbiol 66, 3098-3101. 

A fully automated two-dimensional electrophoresis (2DE) system for rapid and reproducible protein analysis is described. 2DE that is a combination of isoelectric focusing (lEE) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is widely used for protein expression analysis. Here, all the operations are achieved in a shorter time and all the transferring procedures are performed automatically. The system completed the entire process within 1.5 h. A device configuration, operational procedure, and data analysis are described using this system. 

Two-dimensional electrophoresis (2DE) that is a combination of isoelectric focusing (lEF) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) (4) is widely used for protein expression analysis. 

TLC separation of DNP-amino acids may be carried out on cellulose [14] or on polyamide layers [15,16] with several types of solvent systems. A two-dimensional separation of DNP-amino acids on cellulose with toluene—2-chloroethanol—pyridine—5 N ammonia (5 3 1.5 6.5) and saturated ammonium sulfate-water-sodium dodecyl sulfate (25 ml 175 ml 0.144 g) is shown in Fig.4.8. A similar separation on polyamide layers is... 

Macfarlane, D.E. (1989) Two dimensional benzyldimethyl-n-hexadecylaimnonium chloride-sodium dodecyl sulfate preparative polyacrylamide gel electrophoresis a high capacity high resolution technique for the purification of proteins from complex mixtures. Anal. Biochem. 176,457-463. 

This chapter will describe in detail the procedure for Western blotting of polypeptides and proteins separated on a denaturing polyacrylamide gel system. This procedure is used routinely in our laboratory for the analysis of polypeptides from a variety of subcellular fractions of whole tissue and cell lines, and has evolved over a number of years in the hands of several people. Many different immunoblotting procedures are currendy available details of variadons from the I-protein-A method described here are given in the Notes secdon, as are brief amendments covering electrotransfer from two-dimensional and isoelectric focusing systems. A detailed descripdon of sodium dodecyl sulfate polyacrylamide electrophoresis (SDS-PAGE) is not appropriate for this chapter, and the reader is referred to vol. 1, Chapter 6, and refs. 9-14. For details of the producdon of polyclonal and monoclonal andsera, Chapters 1-6 in this vol. 

The combination of CIEF and mass spectrometry is analogous to two-dimensional electrophoresis [4]. In this case, the mass spectrometer provides the molecular-weight information instead of sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This information can be obtained by on-line CIEF-MS [5] or by using CIEF as a micropreparative technique [6]. 

Motta A, Patore A, Goud NA, and Castiglione Morelli MA. Solution Conformation of Salmon Calcitonin in Sodium Dodecyl Sulfate Micelles as Determined by Two-dimensional NMR and Distance Geometry Calculations. Biochemistry 1991 30 10444-10450. 

Two dimensional electrophoresis provide ultrahlgh resolution of various proteins (29). For example, by combining isoelectric focusing (lEF) with perpendicular electrophoresis in the presence of the detergent sodium dodecyl sulfate (SDS), the resulting resolution of 10,000 proteins could be theoretically obtained. Samples of human cells showed 100-2000 protein spots. 

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