Polyacrylamide standards

FIGURE 8.12 Effect of pore diameter on SEC of standards (nondenaturin > mobile phase). Nondenaturing" refers to the effect on the stationary phase. Most iarge proteins were in fact denatured by this mobile phase (which was optimized for use with peptides, not proteins). Accordingly, it was necessary to use polyacrylamide to demonstrate the approximate range and position of Vo under these conditions. The polyacryiamide standards both eiuted at V with the 300-A coiumn (not shown). Columns and flow rate Same as in Fig. 8.11. Mobile phase Same as in Fig. 8.1. Sample key (B) Ovalbumin (43,000 Da) 0) polyacrylamide (1,000,000 Da) (K) polyacrylamide (400,000 IDa) (L) low molecular weight impurity in the polyacrylamide standards. Other samples as in Fig. 8.11. 

FIGURE 9.23 Analysis of ultrahigh poly(acrylamjde). MW 48 million by analytical ultracentrifugation. Eluent 0.1 M Na2SO<. Flow rate 0.3 ml/min. Columns PSS Suprema 30000, 20 /tim, 8 x 300 mm. Oven temp 30°C. Detector Rl. Standards PSS polyacrylamide standards. 

Figure 8. Chromatograms for nonionic polyacrylamide standards and Standard C for the column combination shown in Figure 6.

Estimation of Polymer Sizes by Gel Permeation Chromatography. The copolymer (1 mg) was dissolved in 1 ml of phosphate buffered saline (PBS), pH 7.4, and applied to a column of Sephacryl S-300 (1 X 108 cm) or Sephacryl S-400 (1 x 114 cm). The column was eluted with PBS at a flow rate of 0.2 ml/min. The elution profile of the copolmer was monitored by its absorbance at 214 nm. Bovine serum albumin (BSA) was chromatographed for comparative purposes and polyacrylamide standards (Modchrom, Inc.) were used. 

The Micropak TSK Gel PW, TSK Gel PWXL and Shodex OHpak Q-800, B-800, and KB-800 series are more recently available columns developed for analyzing the acrylamide polymers and other water-soluble polymers in aqueous SEC. The TSK columns have been evaluated by Barth (25), Alfredson et al. (42), Sasaki et al. (43), and Lin and Getman (44). Showa Denko K. K. (45) reported the SEC analysis of PAM by Shodex OHpak columns. The narrow MWD polyacrylamide standards ( =... 

Table 2 Polyacrylamide Standards (Reported by American Polymer Standards Corporation)...

Liquid chromatography at the critical adsorption point (LC LCS) was investigated. The solubility of macromolecules, the sensitivity of LC LCS to such variables as the injection volume, concentration injected, mobile phase flow rate and mixing between the injection zone, were investigated, using polymethyl methacrylate (PMMA) and polyacrylamide standards. It is proposed that using tetrahydrofuran and n-hexane as eluents for PMMA, at the LC LCS, limited polymer insolubility or even local precipitation is combined with the size exclusion of macromolecules. 12 refs. 

Mifflin and associates described a membrane electrode for the quantitative analysis of penicillin in which the enzyme penicillinase is immobilized in a polyacrylamide gel that is coated on a glass pH electrode. The following data were collected for a series of penicillin standards. 

Polyacrylamide, whether charged or not, can be detected by reactions of the amide group (67,68) however, a number of substances can interfere with the determination. If the molecular weight is high enough, flocculation of a standard slurry of clay or other substrate is a sensitive method for detecting low levels of polyacrylamide (69). Once polymers are adsorbed on a surface, many of these methods caimot be used. One exception is the use of a labeled polymer. 

Fig. 3. Sodium dodecyl sulfate—polyacrylamide gel electrophoretic pattern for molecular weight standards (lane 1) water-extractable proteins of defatted soybean meal (lane 2) purified IIS (glycinin) (lane 3) and purified 7S (P-conglycinin) (lane 4) where the numbers represent mol wt x 10. The gel was mn in the presence of 2-mercaptoethanol, resulting in the cleavage of the disulfide bond linking the acidic (A bands) and basic (B bands) polypeptides of the...

Figure 5 SDS Polyacrylamide Gel Electrophoresis of pectinase from different steps of purification A (1,6) standard protein (2) crude enzyme (3)...

Exacting control of buffer preparation and the characteristics of capillaries and coatings is now recognized as key to successful electrophoretic separations.2 Repeatability of separations requires standardized surface preparation and rinse procedures. For example, capillaries can be coated with polyacrylamide using thionyl chloride surface activation. This approach was useful in DNA analysis.3 Non-aqueous buffers can be used to permit the use of thicker capillaries and higher voltages.4... 

The total proteins synthesized by bacteria grown under standard conditions, when analyzed by polyacrylamide gel electrophoresis (PAGE), form patterns that can be compared to those of known strains by visual or computer-assisted... 

Widhalm et al. (1991) reported the use of noncrosslinked polyacrylamide for protein separation in fused silica capillaries. This matrix has low viscosity and can be replaced between separations, greatly facilitating automation of the separation. A wide range of noncrosslinked polymers has been used for size-based protein separations. Noncrosslinked polymers do not form a gel, and it is inappropriate to refer to this separation as gel electrophoresis. A number of names have been used for the method. In an effort to standardize nomenclature, IUPAC has used the term capillary sieving electrophoresis. 

Papermaking technology, 17 496, 497 Paper manufacture. See also Papermaking acrylic ester polymers for, 1 390 polyacrylamide polymers for, 1 324-325 Paper materials standards, 15 742 Paper mills, use of tire-derived fuel in, 21 464... 

RF RIC RMM RSD S/N SDM SD SDS-PAGE spectrometry radiofrequency reconstructed ion chromatogram relative molecular mass relative standard deviation signal-to-noise ratio selected-decomposition monitoring standard deviation sodium dodecyl sulfate-polyacrylamide gel electrophoresis... 

Figure 3 Biosynthesis and purification of 90-kD elastin analogue analyzed by denaturing polyacrylamide gel electrophoresis (10-15% gradient, visualized by silver staining). Lanes 1-7 time course of target protein expression at 0, 30, 60, 90, 120, 150, and 180 minutes after induction. Lane 9 soluble lysate of induced E. coli expression strain BLR(DE3)pRAMl. Lanes 10-13 protein fractions obtained from immobilized metal affinity chromatography of the lysate on nickel-NTA agarose (imidazole gradient elution). Lanes 8,14 protein molecular weight standards of 50, 75, 100, and 150 kD.

Polyacrylic acid (PAA) was obtained from Scientific Polymers, Inc., Ontario, NY, as a secondary standard with a mass-averaged molecular weight of two million. The polyacrylamide (PAM) used was Separan MGL obtained from Dow Chemical Company, Midland, MI. Its reported molecular weight was in the range of 500,000 to 5,000,000. The monomer structures of PAA and PAM are illustrated in Figure 1. 

Figure 6. 10-15% SDS polyacrylamide gel of lane 1 — untreated E, coli extract (0 mM KCl) lane 2 — the peak GUS fraction from the HPLC separation of untreated extract lane 3 — Sigma SDS-7 molecular weight standards (approximate molecular weights of 4 standards are indicated on the right hand edge of the figure) lane 4 — BPA-1000 treated E. coli extract (4000 ppm BPA-1000 and 300 mM KCl) lane 5 — the peak GUS fraction from the HPLC separation of BPA-1000 treated extract. GUS migrates slightly slower than the 66,000 molecular weight standard.

---