SDS polyacrylamide electrophoresis

The following procedure relates to electrophoretic protocols where the first dimension is developed by isoelectric focusing (in tube gels) and the second dimension is a size exclusion separation by SDS polyacrylamide electrophoresis in a slab gel. 

The criterion for purity was a single band on a heavily loaded SDS-polyacrylamide electrophoresis gel. Antibodies were raised in rabbits in response to three intravenous injections each of 2mg. of laccase A with Freunds adjuvant, at 2 weekly intervals. Blood was removed 7d after the last injection and serum collected after coagulation of the blood cells. 

Fig. 6. Kinetics of immobilization of glutaryl-7-ACA-acylase on epoxy-activated polymethacrylate. The Gl-7-ACA-acylase was incubated with the epoxy-activated carrier. At definite times aliquots were taken from the reaction suspension. Supernatant and carrier-fixed enzyme were separated by centrifugation. The carrier-fixed enzyme was washed with water to remove non-covalently linked enzyme. The activities of the immobilized enzyme and supernatant were determined (5 mM potassium phosphate buffer pH 8,37°C, 2% glutaryl-7-amino cepha-losporanic acid, pH-stat 8.0). Simultaneously, an aliquot of carrier-fixed enzyme was boiled in sodium dodecylsulfate (SDS)/glycine buffer and the supernatant was subjected to SDS-polyacrylamide electrophoresis (see insert from left to right lane 1 Carrier-fixed enzyme, 2 h lane 2 Carrier-fixed enzyme, 4 h lane 3 Carrier-fixed enzyme, 6 h lane 4 Carrier-fixed enzyme, 21 h lane 5 Carrier-fixed enzyme, 69 h lane 6 Dialyzed enzyme lane 7 Supernatant, 2 h lane 8 Supernatant, 21 h lane 9 Supernatant, 69 h lane 10 Molecular weight calibration markers)...

Figure 4. Alfalfa somatic embryo proteins separated after extraction by sucrose density gradient separation (left side of chromatogram) and SDS-polyacrylamide electrophoresis (right side of chromatogram). The 2,4-D-treated embryos express more IIS protein as shown in the sucrose gradient. The IIS protein has polypeptides that co-migrate with IIS seed protein as shown by SDS-PAGE. (Reproduced with permission from reference 4. Copyright 1985, Plenum Publishing.)...

Wool is reduced by 0.3 M TGA aqueous solution (pH 11) that contains 8.0 M urea, neutralized to pH 7 by acetic acid, and oxidized by 1.5 M NaBr03 aqueous solution in the presence of a sufficient amount of unreacted TGA. After filtration of the insoluble component, a water soluble protein is obtained [65]. This protein is (1) fix>m amino acid analysis, the SH group that is chemically treated with S-carbox5rmethyl-alanyl disulfide (CMAD —SSCH2COOH) (2) from SDS-polyacrylamide electrophoresis, the mixture of type-1 and type-11 LS keratin proteins and (3) from the CD spectrum, a polymer with a-helix (Fig. 16). This is called carboxymethylalanyl disulfide keratin (CMADK). 

Figure 3. SDS polyacrylamide electrophoresis of ATAa following the second polybuffer exchange column.

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