Micro-Electrophoresis in Continuous-Polyacrylamide-Gradient Gels, II. Fractionation and Dissociation of Sodium Dodecylsulfate Protein Complexes

Separation of aminoacyl‐tRNAPhe and tRNAsPhe with partially hydrolysed 3′‐end by polyacrylamide gradient micro‐electrophoresis

“…The distance between the neighbouring peaks corresponding to the aminoacylated tRNAPhe and the native tRNAPhe in the gel is 130 pm. If the positive charge of phenylalanine is eliminated by the presence of 0.05% sodium dodecyl sulfate in the system [8] the distance between the peak maxima of tRNAPhe-A,, and tRNAPhe-A,, remains unchanged, whereas the distance between native and aminoacyl-tRNA decreases to 100 pm.…”

“…were obtained using the Joyce-Loeble doublebeam microdensitometer with 100: 1 ratio of record to sample travel using a gray wedge of 0.79 d. Further details about the micro-electrophoresis on polyacrylamide gradient gels are presented elsewhere [7,8,9,13].…”

Identification of tubulin associated with rat brain myelin

“…10% glycerol/5% 2-mercaptoethanol/l% SDS/O.0625 M Tris-HCl (pH 6.8)/0.001% bromophenol blue, and heated at 1OO'C for 5 min. Samples were subjected either to l-4% gradient microgels as in [7], or 0.75 mm thick IO-2% gradient slab gels using a discontinuous SDS-containing buffer system [22,23]. For quantitative evaluation, the photographic transparencies of the Coomassie brilliant blue stained gels were scanned at 560 nm in a Gilford 2400-S spectrophotometer.…”

“…The myelin fractions were analyzed by electro-170 phoresis on microgels [7] for the presence of polypeptides with the same mobility as tubulin. When a DEAE-cellulose purified rat brain tubulin marker was added to a myelin fraction, enhancement of a pre-existing band was apparent, suggesting that tubulin is one of the constituents of the myelin ( fig.…”

Posttranslational Folding of α1-Inhibitor 3

“…The cleavage products were then detected by SDS-PAGE under reducing conditions. Since artifactual bands have been reported to occur with this technique, the results should be treated with some caution (Rü chel et al, 1974). With pulse-labeled ␣ 1 I 3 ( Fig.…”