Gradient gels linear

Fig. 6.3. SDS/PAGE (5-15% linear gradient gel) separated proteins of the total worm homogenate and isolated brush border fractions from protoscoleces of Echinococcus granulosus (horse strain), (a) Coomassie blue staining (b) Periodic acid-Schiff (PAS) staining. (After McManus Barrett, 1985.)...

Noda, K., Kubota, K., Yamasaki, R. Separation of lipooligosaccharides by linear gradient gel electrophoresis. Anal Biochem 279 (2000) 18-22. 

Figure 9.10. Voltage versus distance for (a) a linear electrophoresis gel, and (b) a gradient gel. From Ohm s law, with constant current, the observed voltage changes mirror the resistance properties of the gels.

Figure 9.11. Comparison of the band separation of linear, wedge, and ionic strength gradient gels. The T reaction of a sequencing experiment carried out on M13 mp8 DNA was run on three different 40 cm 6% polyacrylamide gels, using (a) a standard linear gel, (b) a 0.35-1.05-mm wedge gel, and (c) a 0.05 0.50 Af Tris buffer gradient. [Reprinted, with permission, from A. T. Bankier and B. G. Barrell, in Nucleic Acids Sequencing A Practical Approach , C. J. Howe and E. S. Ward, Eds., Oxford University Press, New York, 1989. IRL Press at Oxford University Press 1989.]...

Figure 11.6. Experimental arrangement for casting an immobilized pH gradient gel. A linear pH gradient is generated by mixing two solutions with initially equal volumes, where one solution is dense and the other is, and they are titrated to the extremes of the desired pH interval. The compensating rod in the reservoir is used as a stirrer after the addition of catalysts and for hydrostatically equilibrating the two solutions.

The most frequently used version is that with a linear gradient gel that can be prepared by mixing two differently concentrated gel solutions in two connected jars [104]. Thus, for a 4-26% gel gradient the following solutions are recommended (ammonium persulphate is added to the mixture immediately before mixing is started). 

Fig. 3A,B. SDS gradient gel electrophoresis and autoradiography of ADP-ribosylated proteins. Permeabilized HeLa cells (5 X 10 ) were incubated for 20 min at 26°C with 0.133 ijM [ P]-NAD, sp.act. 245.8 Ci mmoP, made up to 1 nM or 100 /lAf with non-radioactive NAD. Each assay contained about 5.9 X 10 cpm. Reactions (75 n ) were stopped by adding 75 n of solution containing 2% (w/v) SDS, 50 mM 2-mercaptoethanol, 6 M urea, 10 mM sodium bisulphite, 5 mM 4-aminobenzamidine, 0.5 mM PMSF, 0.15 iig of pepstatin and 0.15 Mg of leupeptin in 100 mM Tris-HCl pH 5.4 and 40% (v/v) glycerol. The mixture was immediately boiled for 4 min. Slab-gel electrophoresis was carried out in a 6-18% linear gradient polyacrylamide gel made in 25 mM sodium phosphate pH 6.8,1% (w/v) SDS and 3 M urea. A 4% stacking gel was used made in 10 mM sodium phosphate pH 6.0. The electrode buffer was 25 mM sodium phosphate pH 6.8, 1% SDS. Gels were subjected to electrophoresis for 6-7 h at a constant current of 40 mA.

Figure 14.7 Concentration gradients (assumed linear) resulting from the boundary conditions in Turing pattern experiments with the CDIMA reaction. The position variable has been normalized so that the length of the gel is 1. (Adapted from Lengyel et al., 1992a.)...

Fig. 10. HPLC of proteins (commercial samples) on the /V-butyl polyacrylamide coated silica gel column. Sample 20 pi of 5-15 mg/ml protein solution in buffer A. Buffer A 10% methanol, 0.2 mol/1 ammonium acetate, pH 4.5. Buffer B methanol. Gradient 50-min linear, 0-100% B. Flow rate 0.8 ml/min. Peaks (/) — lysozym, (2,3) — insulin, (4,5) — myoglobin [57]...

The sample is loaded at a flow-rate of 1 ml/min onto the FPLC column equilibrated with the same MOPS buffer used to resuspend the RNA pellets. The free nucleotides are completely removed with a 5-ml wash with 350 mM NaCl and the RNA is eluted with a 20-ml (350—750 mM NaCl) linear gradient and analyzed by PAGE/urea gel electrophoresis (see later). Up to 2 mg of RNA can be loaded onto and eluted from a 1-ml (of resin) mono Q column without loss of resolution. The homogeneity of RNA in the fractions collected, as seen by gel electrophoresis, should be >90%. The appropriate fractions are pooled and the RNA collected by ethanol precipitation. The RNA pellet is washed twice with 70% ethanol, air-dried, and finally redissolved in DEPC-treated H20. The total recovery after the entire procedure of purification is = 90%. This protocol yields = 800 pmoles of purified 002 mRNA/pmole template DNA. 

Fig. 12. Separation of styrene oligomers by reversed-phase (left) and size-exclusion chromatography (right) (Reprinted with permission from [121]. Copyright 1996 American Chemical Society). Conditions (left) column, molded poly(styrene-co-divinylbenzene) monolith, 50 mm x 8 mm i.d., mobile phase, linear gradient from 60 to 30% water in tetrahydrofuran within 20 min, flow rate 1 ml/min, injection volume 20 pi UV detection, 254 nm (right) series of four 300 mm x 7.5 mm i.d. PL Gel columns (100 A, 500 A, 105 A, and Mixed C), mobile phase tetrahydrofuran, flow rate, 1 ml/min injection volume 100 pi, toluene added as a flow marker, UV detection, 254 nm temperature 25 °C,peak numbers correspond to the number of styrene units in the oligomers...

Figure 3.15 Chromatogram of fibre-type proteins on polystyrene gels having different pore sizes. Column A, PLRP-S 300 A, 15 cm x 4.6 mm i.d. B, PLRP-S 1000 A (polystyrene gel), 15 cm x 4.6 mm i.d. eluent, 15 min linear gradient from 20% of 0.25% trifluoroacetic acid to 60% of 0.25% trifluoro-acetic acid in 95% aqueous acetonitrile flow rate, 1.0 ml min-1 detection, UV220 nm. Peaks 1, collagen (Mr 120 000) and 2, fibrinogen (Mr 340 000). (Reproduced by permission from Polymer Laboratories data)...

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