SDS acrylamide gel electrophoresis

A few practical notes should be emphasized regarding SDS electrophoresis. Potassium salts should be avoided and sodium salts used in their places because potassium dodecyl sulfate is quite insoluble. In addition, even sodium dodecyl sulfate is insoluble below about 10°C. Although not specifically cited above, disc gel electrophoresis may also be used in the presence of SDS. These techniques, described by Studier (11) and Ames (12), are of great advantage when the sample volume is in excess of 10 to 20 /Ltliters per gel. A widely used buffer system for SDS acrylamide gel electrophoresis is that developed by Laemmli (41). 

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Fig. 2.4. Vector characterization. (A). SDS acrylamide gel electrophoresis of a rAAV-UF-11 vector. Approximately 2.5 x 1011 drp were loaded on a 10% SDS-polyacrylamide gel. The three capsid protein bands in the correct stoichiometry are visualized by silver staining. (B) Infectious center assay of a rAAV-CB-hAAT stock when probed for the transgene each spot represents a vector infected cell. (C) Dot blot analysis of the rAAV stock to determine the titer of DNase-resistant particles of rAAV. Amounts (ng) refer to a serial dilution of the packaged rAAV vector plasmid used to construct the standard curve. The stock has a calculated titer of 4.3 x 1013 drp/ml.

Figure 5. SDS-acrylamide gel electrophoresis of brush border membrane proteins from rats of adequate zinc status (0), following a 4-day zinc depletion (4), and following an 8-day zinc depletion (8). Proteins migrating with approximate molecular weights of 63,000 and 45,000 daltons are more apparent after 4 and 8 days of dietary zinc restriction.

II,A,3 SDS-Acrylamide gel electrophoresis of the soluble NADH dehydrogenase derived from complex I has shown that this enzyme is composed of two subunits with molecular weights of approximately 28,0CX) and 56,000 (G. Dooijewaard and E. C. Slater, private communication). 

Figure 6-14. Cross-section of an isoelectric focusing gel on top of a slab gel in preparation for SDS acrylamide gel electrophoresis of proteins previously separated by isoelectric focusing. [From P. H. O Farrell, J. Biol. Chem., 250 4007 (1975).]...

Figure 6-20. Polio virus-induced proteins from the cytoplasm of infected HeLa cells. Proteins were separated by SDS acrylamide gel electrophoresis and the gels fractionated using an autogeldivider. (O) Distribution of tritium-containing proteins from cells infected and then labeled under conditions where only virus-specified proteins are made. ( ) Distribution of the label of C labeled purified virions. [From Summers et al., Proc. Natl. Acad. Sci. US, 54 505 (1965).]...

During the 4 week period used to obtain control sera, determine the purity of a commercial avidin preparation using SDS-acrylamide gel electrophoresis as described in Chapter 5. Use only preparations exhibiting a single electrophoretic species in these experiments. 

Figure 6 shows the determination of the subunit size of transketolase by SDS acrylamide gel electrophoresis. 

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