Polyacrylamide activation

Low molecular weight (1000—5000) polyacrylates and copolymers of acryflc acid and AMPS are used as dispersants for weighted water-base muds (64). These materials, 40—50% of which is the active polymer, are usually provided in a Hquid form. They are particularly useful where high temperatures are encountered or in muds, which derive most of their viscosity from fine drill soHds, and polymers such as xanthan gum and polyacrylamide. Another high temperature polymer, a sulfonated styrene maleic—anhydride copolymer, is provided in powdered form (65,66). AH of these materials are used in relatively low (ca 0.2—0.7 kg/m (0.5—2 lb /bbl)) concentrations in the mud. 

Coagulation involves the addition of chemicals to alter the physical state of dissolved and suspended solids. This facilitates their removal by sedimentation and filtration. The most common primary coagulants are alum ferric sulfate and ferric chloride. Additional chemicals that may be added to enhance coagulation include activate silica, a complex silicate made from sodium silicate, and charged organic molecules called polyelectrolytes, which include large-molecular-weight polyacrylamides, dimethyl-diallylammonium chloride, polyamines, and starch. 

J. N. Kinkel, Optically active polyacrylamide/silica composites and related packings and their application to chiral separations in A practical approach to chiral separations by liquid chro-matogratphy, G. Subramanian, VCH, Weinheim (1994) Chapter 8. 

To develop a continuous process, the immobilisation of aminoacylase of Aspergillus oryzae by a variety of methods was studied, for example ionic binding to DEAE-Sephadex, covalent binding to iodo-acetyl cellulose and entrapment in polyacrylamide gel. Ionic binding to DEAE-Sephadex was chosen because the method of preparation was easy, activity was high and stable, and regeneration was possible. 

The described bioaffinity separations demonstrate that polyacrylamide spacers aid the selective binding of highly complex and delicate biomacromolecules and their associates. Moreover, these solutes remain biologically active after desorption probably due to the high inertness and flexibility of the surrounding polymer chains fixed on the solid support. The unbound parts of serum usually show no loss of the activities of their constituents. Thus we evaluate the surface of inorganic supports coated with chemisorbed iV-hydroxyethyl polyacrylamide and its derivatives as being biocompatible. 

Mannich polymers Cationic, solution polymer flocculants with a MW of 5 to 8 M. Very high viscosity at only 4 to 8% active strength solution liquid. A hydrolyed polyacrylamide, it is very useful for general municipal waste water treatment. The dose rate normally is 200 to 300 ppm or more. 

Polyacrylamide Nonionic flocculant, usually supplied as 100% active powders but also available as 30% or so emulsions. MW ranges typically from 2.5 M to 20 M. Test for suitability using jar-tester apparatus at, say, 1 to 5 ppm. It can be used as a coagulant aid for alum, and PWG are available. 

A wide range of polyacrylic acids (PAAs), polymethacrylic acids (PMAAs), polyacrylamides (PAMs), their salts and homo-, co-, and terpolymer derivatives are available from a great many manufacturers around the world today. Polyacrylates especially are a backbone of BW chemical formulations and are manufactured with an almost infinite variety of average molecular weights (MWs), MW distributions, activity strengths, and other characteristics. 

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. 

Figure 5. Analytical isoelectric focusing. Ultrathin layers (0.4 nun) of polyacrylamide with ampholytes pH 2-11 were used. Samples of 10 pg of protein in 10 pi of 1 % glycine were applied. A.- Silver staining. B.- Stain for activity on overlays containing pectin in tris/HCl buffer at pH 8.0 with CaClj M.- Broad pi Calibration Kit protein (Pharmacia), samples of 5 pg of protein were applied. 1.-Ammonium sulphate precipitated proteins from cultures on pectin. 2.- Fractions with PNL activity eluted from the Superdex 75HR1030 column. 3.- Purified PNL.

Fig. 5. Isoelectric focusing (pH gradient 3-10) of Fraction A and Fraction B in ultrathin polyacrylamide layers. 5 pg of fractions were applied. Activity detection with ruthenium red (left) and with Ostazin Brilliant Red/D-galacturonan DP 10 agar print (right).

Ultrathin-layer isoelectric focusing in polyacrylamide gels on polyester films was performed as described (Radola, 1980). Polygalacturonase activity was detected by the print technique with a dyed substrate (Ostazin Brilliant Red-D-galacturonan DP 10) (Markovic et al., 1992) or by the print technique with colouress D-galacturonan DP 10 dyed additionally with ruthenium red (Sigma, Germany). 

Fig. 3. Isoelectric focusing in ultrathin polyacrylamide layers (pH gradient 3 -10) of multiple forms of polygalacturonase produced by Candida boidinii under different cultivation conditions a - pectin, pH 3.51 b - pectin, pH 5.49 c -pectin, pH 7.01 d - 20% of D-galactopyranuronic acid in pectin e - pectate. A - Activity detection with print technique on colouress D-galacturonan DP 10 dyed additionally with ruthenium red ( both exo- and polygalacturonases) and B - activity detection with Ostazin Brilliant Red/D-galacturonan DP 10 agar print (polygalacturonases).

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.

Assays. Protein concentrations were measured by the method of Bradford (18) and the various contractile protein ATPase activities by tRe method of Martin and Doty (19). Gel electrophoresis was carried out by the method of Ames (20) on 1.5 ran polyacrylamide slabs using the discontinuous SDS buffer system of Laemnli (21). Dried gels were scanned at 550 nm for densiometry measurements. 

The entrapment method is based on confining the enzyme within the lattice of a polymeric matrix. Polyacrylamide gels have successfully yielded stable enzyme films with a high retention of activity... 

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... 

Another approach has been to immobilize proteins within arrays of microfabricated polyacrylamide gel pads (Arenkov et al., 2000). Nanoliters of protein solutions are transferred to 100 x 100 x 20-pM gel pads and assayed with antibodies that are labeled with a fluorescent tag. Antigen imbedded in the gel pads can be detected with high sensitivity and specificity (Arenkov et al., 2000). Furthermore, enzymes such as alkaline phosphatase can be immobilized in the gel pads and enzymatic activity is readily detected upon the addition of an indicator substrate. The main advantage of the use of the threedimensional gel pad for fixation of proteins is the large capacity for immobilized molecules. In addition, the pads in the array are separated from one another by a hydrophobic surface. Thus, each pad behaves as a small test tube for assay of protein-protein interactions and enzymatic reactions (Arenkov et al., 2000). The disadvantage of the method is the need to microfabricate the array of gel pads in that microfabrication is... 

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