Polyethylene glycol electrophoresis

Charlionet, R Levasseur, L Malandain, 11, Eliciting Macroporosity in Polyacrylamide and Agarose Gels with Polyethylene Glycol, Electrophoresis 17, 58, 1996. 

In the previously described electrophoretic methods, the capillary was filled with electrolytes only. Another mode of operation in capillary electrophoresis involves filling the capillary with gel or viscous polymer solutions. If desired, a column can be packed with particles and equipped with a frit.68 This mode of analysis has been favorably used for the size determination of biologically important polymers, such as DNA, proteins, and polysaccharides. The most frequently used polymers in capillary gel electrophoresis are cross-linked or linear polyacrylamide,69 cellulose derivatives,70-75 agarose,76 78 and polyethylene glycols. 

Ceruloplasmin (from human blood plasma) [9031-37-2]. Purified by precipitation with polyethylene glycol 4000, batchwise adsorption and elution from QAE-Sephadex, and gradient elution from DEAE-Sepharose CL-6B. Ceruloplasmin was purified 1640-fold. Homogeneous on anionic polyacrylamide gel electrophoresis (PAGE), SDS-PAGE, isoelectric focusing and low speed equilibrium centrifugation. [Oestnuizen AB 146 1 1985]. 

Figure 6. Results of SDS polyacrylamide gel electrophoresis on the supernatant of a crude maize extract (A), a 6 to 14 % polyethylene glycol precipitate (B), and pooled fractions having ACCase activity from a Sephacryl S-300 column (C).

Upon electrophoresis, macro-AMY usually forms a broad migrating band, clearly different from the homogeneous bands that are produced by AMY isoenzymes present in serum (Figure 21-6). If electrophoretic separation is not available, precipitation of the macrocomplex by a polyethylene glycol (PEG) 6000 solution (240 g/L) represents a good alternative. Residual AMY activity of less than 30% in the supernatant is indicative of macroamylasemia. ... 

Extracts of the R and S biotypes of Eleusine were fractionated by stepwise increases in polyethylene glycol concentration and the various fractions were monitored for the presence of tubulin and other proteins by electrophoresis and Western blotting. By making small increases in PEG concentration, one fraction contained virtually all the recognizable tubulin and was > 85% pure as determined by Coomassie blue staining of denaturing gels (23, 24). This is comparable in purity to the protocols of Morejohn gi al- (25), but is a much faster method and allows... 

Cunico R L, Gruhn V, Kresin L, et al. (1991). Characterization of polyethylene glycol modified proteins using charge-reversed capillary electrophoresis. /. Chromatog. 559 467-477. 

Li W, Zhong Y, Lin B, Su Z (2001). Characterization of polyethylene glycol-modified proteins by semi-aqueous capillary electrophoresis. J. Chromatog. A. 905 299-307. 

Nagata et al. reported the use of a polyethylene glycol-coated poly(methyl methacrylate) (PMMA) microchip for microchip gel electrophoresis separations. Trypsin inhibitor, bovine serum albumin (BSA) and 3-galactosidase (labeled with Alexa Ruor 488) were electrokinetically loaded for 60 s. The sodium dodecyl sulfate (SDS)-protein complexes were then separated and baseline resolved in 8 s using a 5% linear polyacrylamide/0.1% (w/v) SDS-based separation buffer, 3 mm separation channel, and 303 V cm electric field. 

Nagata, H., Tabuchi, M., Hirano, K., and Baba, Y. High-speed separation of proteins by microchip electrophoresis using a polyethylene glycol-coated plastic chip with a sodium dodecyl sulfate-hnear polyacrylamide solution. Electrophoresis 26, 2687, 2005. 

Figure 2 Serum electrophoresis of a patient with a small monoclonal band (top) CE (bottom) agarose electrophoresis l = internal standard A = albumin ai =ai-globulin a2 = a2-glob-ulin, T = transferrin C = complement M = monoclonal band > = >-globulins X = marker). CE conditions capillary 30cmx 50 pm (ID), 9kV, detection at214nm. Separation buffer 7g boric acid, 7g sodium carbonate, and 5 g of polyethylene glycol 8000 in 1000 ml water.

Fig. 5 Extraction of analytes with organic solvents (A) Benzoic acid standard 30 mg/L in the electrophoresis buffer (separation under non-stacking) and (B) Benzoic acid 30 mg/L in 10 mmol/L HCl extracted with chloroform (containing 10% methanol) with direct chloroform injection for 40 sec on a capillary 50 jjim x 40 cm, 214 nm, and 12 kV. Separation buffer boric acid 400 mg, sodium carbonate 200 mg, polyethylene glycol 200 mg in 100 ml water. (B = benzoic acid.)...

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