Acrylamide electrophoresis

Figure 3-10 Estimation of the molecular mass of the polypeptide chain of the nitrogenase Fe-protein using SDS-poly-acrylamide electrophoresis from a set of four standard curves. The marker proteins are (1) catalase, (2) fumarase, (3) aldolase, (4) glyceraldehyde-phosphate dehydrogenase, (5) a-chymotrypsinogen A, and (6) myoglobin, (o) indicates position of azoferredoxin. From Nakos and Mortenson.195...

Fig. 8,7. Routine for two-dimensional acrylamide electrophoresis, from Vigne and Jordan (1971) (for explanation see text). Other workers prefer to embed the gel strip after the first separation at the bottom of the slab, for the samples to migrate upwards...

Vertical acrylamide electrophoresis unit with power supply (Bio-Rad). 

Human serum 371,000 Sepharose 6B gel filtration acrylamide electrophoresis PIO... 

Song, L., Liang, D., Kielescawa, J., Liang, J., Tjoe, E., Fang, D., and Chu, B., DNA sequencing by capillary electrophoresis using copolymers of acrylamide and Af,Af-dimethyl-acrylamide. Electrophoresis 2001, 22, 729-736. 

Molded polyamide surfaces can be hardened by grafting with Ai,Ai-diallylacrylamide [3085-68-5] monomer under exposure to electron beam (159). AijAZ-DiaHyltartardiamide [58477-85-3] is a cross-linking agent for acrylamide reversible gels in electrophoresis. Such gels can be dissolved by a dilute periodic acid solution in order to recover protein fractions. 

Polyacrylamide Electrophoresis. Polyacrylamide gels are synthesized through the combination of acrylamide [79-60-1] (qv), CH2=CHC0NH2, monomer and a cross-linking comonomer (see Acrylamide POLYMERS). Typically, the cross-linking comonomer of choice is... 

Raymond, S Weintranb, LS, Acrylamide Gel as a Supporting Medium for Zone Electrophoresis, Science 130, 7111,1959. 

Figure 3. SDS-PAGE and in situ pectinase activity on pectin and polygalacturonic acid-agarose overlays of culture filtrates of Aspergillus niger N-402 (upper panel) and Aspergillus FP-180 (lower panel) at 2.5, 3.5, 5.5 and 6.5 pHi (Lanes a, b, c, and d, respectively). Electrophoresis on 10% acrylamide slab gel (14 X 8 cm) in the presence of SDS was according to Laemmli (6), run at 30 mA constant current for 2 hours. Crude cell-free samples were concentrated by lyophilization, dialyzed, boiled with sample buffer by 60 sec. and applied to each well. Polyacrylamide gel and overlays were incubated overnight with 0.17 acetate buffer at room temperature.

Raymond, S. and Weintraub, L., Acrylamide as a supporting medium for zone electrophoresis, Science, 130, 711, 1959. 

Analysis of the mRNA quality by PAGE/urea It is good practice to check the quality of the transcription product at various stages of the preparation. For this purpose, aliquots (e.g., 5 pi) of the reaction mix are taken at the beginning and end of the transcription reaction as well as after each step of the purification and mixed with an equal volume of electrophoresis sample buffer. After incubation at 65° for 5 min, the samples are loaded on a 6 to 8% acrylamide-7-8 M urea gel. 

Preparative electrophoresis on Sephadex G-25 (Ref. 168) or double isoelectric focusing,208 preceded by chromatography on Sephadex G-75, CM-cellulose, and calcium phosphate, was used for the isolation of endo-D-galacturonanase from the filtrate of a Verticillium albo-atrum culture. The homogeneity was confirmed in both cases by electrophoresis on poly(acrylamide) gel. The molecular weight of the enzyme was close to the values found for Aspergillus endo-D-galacturonanases. 

Figure 7.1. Effect of GM-CSF on neutrophil protein biosynthesis. Human neutrophils were incubated in RPMI1640 medium supplemented with 2.5% foetal calf serum and 60 jtCi/ml [35S]-methionine, in the presence and absence of 50 U/ml GM-CSF. After 4 h incubation at 37 °C, the cells were pelleted by low-speed centrifugation. The proteins in the cell pellets were precipitated by 10% trichloracetic acid and then analysed by two-dimensional polyacrylamide gel electrophoresis, using iso-electrofocussing in the first dimension. Electrophoresis in the second dimension was performed in the presence of SDS and used a 12% acrylamide gel. Source Experiment of Becky Stringer.

Polyacrylamide gel is the most commonly used type of support medium for gel electrophoresis, and polyacrylamide gel electrophoresis is simply known as PAGE. The gel is usually formed by polymerization of acrylamide and the cross-linking agent N, iV -methylene-bis-acrylamide (Bis) in the presence of ammonium persulfate (APS, initiator) and N, N, N, iV -tetramethyl-ethylenediamine (TEMED, accelerator). The total concentration of acrylamide... 

CEC capillary columns filled with hydrophilic polymer gels mimic those used for capillary gel electrophoresis [91]. Typically, the capillary is filled with an aqueous polymerization mixture that contains monovinyl and divinyl (crosslinking) acrylamide-based monomers as well as a redox free radical initiating system, such as ammonium peroxodisulfate and tetramethylethylenediamine (TEMED). Since initiation of the polymerization process begins immediately upon mixing all of the components at room temperature, the reaction mixture must be used immediately. It should be noted, that these gels are very loose, highly swollen materials that usually contain no more than 5% solid polymer. 

Polyacrylamide shows many advantages over starch gel as a medium for high resolution electrophoresis and because of its synthetic nature its pore size can be more easily controlled. The gel is formed by the polymerization of the two monomers, acrylamide and a cross-linking agent, N, iV-methylene-bis-acrylamide (Figure 3.26). The proportion of the two monomers and not their total concentration is the major factor in determining the pore size, the latter having more effect on the elasticity and... 

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