PAGE analysis

For the purification of RHG, which has been described before (6), A. aculeatus was grown in sugar-beet pulp medium for 48 h. From a 2 1 culture containing 3.2 g protein, about 5 mg RHG was obtained after purification. Analysis by SDS-PAGE revealed only one band of about 55 kDa and thus the enzyme appeared to be pure. After N-glycanase treatment and SDS-PAGE analysis, a smaller protein band of about 46 kDa was visible (Fig.l). 

Compounds not eliminated by the counterscreen are re-tested in a secondary assay involving in vitro translation of FF/HCV/Ren mRNA in Krebs-2 extracts using 35S-methionine followed by SDS-PAGE analysis to monitor protein synthesis... 

EFFECT OF SAMPLE PREPARATION ON CE-SDS AND SDS-PAGE ANALYSIS OF RMABS... 

SDS-PAGE analysis of purified FDH revealed two subunits and, in combination with native PAGE, suggested a a2 i structure. Selenium was not released from the protein during denaturing gel electrophoresis and was... 

Cerfain proteins were confirmed by Western blot analysis. The 2D PAGE analysis can resolve up to 2000 proteins. In this study, 820 spots were mapped to proteins but only 165 were used in the protein-to-gene analysis. Presumably these represent the highest fold expressed genes from the Ixmg tumors that were also visible on the gels by silver staining. 

Fig. 2.2.4.1 SDS-PAGE analysis of recombinant LK-ADH purification. Lane 1 molecular weight standards (kDa) Lane 2 crude extract of recombinant . coli BL21(DE3)/pADH Lane 3 ADH after anion-exchange chromatography.

In order to investigate whether tomatinases from F. oxysporum and F. solani share similar molecular characteristics, F. solani tomatinase was partially purified. Comparative SDS-PAGE analysis of the protein fractions with and without tomatinase activity showed the presence of a 32.5 kDa band in all positive fractions, while this band was absent in fractions without tomatinase activity The apparent molecular mass of tomatinase of F. solani differs from that of F. oxysporum (50 kDa), S. lycopersici (110 kDa) [33], and Botrytis cinerea (70 kDa)[36]. The F. solani tomatinase presents a very low activity compared with F. oxysporum enzyme [35, 89]. Western blot analysis showed that the two enzymes also differ in their immunological characteristics since the polyclonal antibody against tomatinase of F. oxysporum f. sp. lycopersici did not recognize the tomatinase from F. solani. These results suggest that the enzyme from F. solani is a novel tomatinase species. 

During this 30 min period, prepare and warm a gel for 6% denaturing PAGE analysis. 

Figure 8. Effect of UV light on binding of [3H]-CPZ to canine striatal homogenates. Fluorogram of irradiated (left) and non-irradiated samples (right) after SDS-PAGE analysis. Taken from the Ph. D. Thesis of K. Thermos.

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