Non-denaturing polyacrylamide gel

Middle panel Cell wall proteins were isolated, 10 pgm of each resolved by non-denaturing polyacrylamide gel electrophoresis and PGl and PG2 isoforms detected by activity staining. 

Its relative electrophoretic mobility in denaturing and non-denaturing polyacrylamide gels indicated that it had a dimeric structure with an estimated molecular mass of 65kDa per monomer (Figure 11.1). This was confirmed by... 

F. oxysporum f. sp. melonis, which was resolved by non-denaturing polyacrylamide gel electrophoresis into active bands having molecular weights of 170-880 kDa with increments of 70 kDa, that is into protein species made of four to 26 subunits... 

Components of a Non-Denaturing Polyacrylamide Cel A non-denaturing polyacrylamide gel is composed of the following components ... 

Preparing a Non-Denaturing Polyacrylamide Gel Electrophoresis with an 8% nondenaturing polyacrylamide gel is commonly used and offers fine separation with high resolution of DNA fragments. Table 3.10 includes the components and the quantities to prepare an 8% non-denaturing polyacrylamide gel. 

The DNA-porphyrin strands 32 were then mixed with 4 equivalents of a complementary 20-mer non-porphyrinic strand, resulting in a doublehelical complex with two DNA-porphyrin strands linked to four complementary strands 35. The stoichiometry was confirmed by absorption spectroscopy and non-denaturing polyacrylamide gel electrophoresis. 

Single-strand conformation polymorphism was originally described by Orita et al. [94] for the detection of point mutations. The idea of SSCP is to perform electrophoresis on a non-denaturing polyacrylamide gel using small PCR products after denaturation of the DNA. As the PCR product moves through the gel, it will regain a secondary structure that is sequence dependent (similar to RNA secondary structure). The mobility of the single-stranded PCR products depends on their secondary structure. Therefore, PCR products that possess sequence differences (mutations, insertions or deletions) will have different mobilities. 

FNR is distributed between the stroma and the thylakoid membrane of higher plants. Estimates of the proportion bound to the spinach thylakoid membrane vary from 40 to 80% (3,4). FNR is probably attached to the membrane by a specific binding protein of 17 kDa (5). Both stromal and bound forms often show considerable heterogeneity when analysed by SDS and non-denaturing polyacrylamide gel electrophoresis. Indeed, up to 8 forms of spinach FNR have been separated by isoelectric focussing(6). The significance, if any, of these variants is not yet clear. 

FIGURE 2. Expression of the pea FNR gene in transgenic tobacco plants. NADPH -nitro blue tetrazolium reductase activity was detected after non-denaturing polyacrylamide gel electrophoresis of leaf extracts from pea (P), transgenic tobacco (Tr) and wild-type tobacco (T). 

Fig. 2. Purified mushroom PPO fractions run on a non-denaturing polyacrylamide gel (10% PAGE). Lanes 1 and 9 are blanks where sample buffer only was applied lanes 2, 5 and 3, 6 correspond to aliquots of peaks A and B of the Igg-Sepharose column, respectively. Lanes 4 and 7 are of unpurlfled enzyme and lane 8 is an aliquot of redlssolved freeze dried unpurlfled enzyme (Sigma Chemical Co.). The left part of the gel (lanes 1, 2, 3 and 4) was stained by substrate (DL-dopa 5 nM) reaction In 0.1 sodium phosphate buffer, pH 7.0, with oxygen replenished by bubbling through the Incubation solution. The band In zone a stained brown immediately, while there was a delay of 10 min In the start of color appearance In zone b. The right hand side of the gel (lanes 5, 6, 7, 8, 9) was stained for protein with Coomassle brilliant blue.

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