Gel , disruption

Finally, an important feature of gels made of adhesive emulsions arises from the deformation of the droplets. Indeed, as the temperature is lowered the contact angles between the droplets increase [27,28] (see Chapter 2, Section 2.3). Consequently, the structure of the final floes depends on the time evolution of the strength of the adhesion. Initially, the adhesion results in the formation of a random, solid gel network in the emulsion. Further increase of adhesion causes massive fracturing of the gel, disrupting the rigidity of the structure and leading to well separated, and more compact floes [27,28]. 

The rheology of the sol-gel transition was undertaken with special care in order to avoid gel disruption. A critical behaviour for the shear modulus with respect to the helix amount, is noticed. A simple relation between the rheological parameters and the degree of helix formation is pointed out in a limited range of helix amounts (X<15%). These experiments will continue on the fully matured gels. 

If, however, quantitation via gel disruption is foreseen, it is advisable to replace the Bis cross-linking reaction with another one that yields more easily solubilizable gels. Ethylenediacrylate is used for this purpose [253]. Dissolution can be done either by the action of concentrated NH4OH [254] or by piperidine-alcoholic hyamine [253] (9 1). In the latter case gels are dissolved by incubation at 37°C for 3-4 h. Dissolution can be accelerated by addition of another portion of piperidine. 

Product recoveiy from reversed micellar solutions can often be attained by simple back extrac tion, by contacting with an aqueous solution having salt concentration and pH that disfavors protein solu-bihzation, but this is not always a reliable method. Addition of cosolvents such as ethyl acetate or alcohols can lead to a disruption of the micelles and expulsion of the protein species, but this may also lead to protein denaturation. These additives must be removed by distillation, for example, to enable reconstitution of the micellar phase. Temperature increases can similarly lead to product release as a concentrated aqueous solution. Removal of the water from the reversed micelles by molecular sieves or sihca gel has also been found to cause a precipitation of the protein from the organic phase. 

Superdex and prepacked Superdex columns are supplied in 20% ethanol. All Superdex may be autoclaved repeatedly at pH 7, 120°C without significant changes in porosity or rigidity. Freezing and thawing of Superdex-based gels may result in disruption of the bead structure and should be avoided. 

It is well known that lyophilic sols are coagulated by the removal of a stabilizing hydration region. In this case, conversion of a sol to a gel occurs when bound cations destroy the hydration regions about the polyanion, and solvated ion-pairs are converted into contact ion-pairs. Desolvation depends on the degree of ionization, a, of the polyacid, and the nature of the cation. Ba ions form contact ion-pairs and precipitate PAA when a is low (0-25), whereas the strongly hydrated Mg + ion disrupts the hydration region only when a > 0-60. 

Protein recovery via disruption has also been achieved by adsorbing water from the w/o-ME solution, which causes protein to precipitate out of solution. Methods of water removal include adsorption using silica gel [73,151], molecular sieves [152], or salt crystals [58,163], or formation of clanthrate hydrates [154]. In most of the cases reported, the released protein appeared as a solid phase that, importantly, was virtually surfactant-free. In contrast to the dilution technique, it appears that dehydration more successfully released biomolecules that are hydrophilic rather than hydrophobic. 

This method can, in principle, be used to determine the transport characteristics of drugs dissolved or suspended in pharmaceutical gels. A potential problem with this method is the possibility for the gel to dissolve into the aqueous phase, with resulting disruption of the free boundary. These changing experimental conditions would lead to lack of precision for the experimental results and difficulty in interpreting them. 

Type 1 gels are mesophases that are so highly ordered that they resist disruption of their structure and are thus extraordinarily viscous, to the point of appearing solid-like, even though no high molecular weight species need be present in the system. Surfactants, both synthetic (e.g., sodium dodecylsulfate) and natural (e.g., phospholipids), and clays are typical representatives of this class. 

Figure 7.1 Separation of proteins by SDS-PAGE. Protein samples are incubated with SDS (as well as reducing agents, which disrupt disulfide linkages). The electric field is applied across the gel after the protein samples to be analysed are loaded into the gel wells. The rate of protein migration towards the anode is dependent upon protein size. After electrophoresis is complete, individual protein bands may be visualized by staining with a protein-binding dye (a). If one well is loaded with a mixture of proteins, each of known molecular mass, a standard curve relating distance migrated to molecular mass can be constructed (b). This allows estimation of the molecular mass of the purified protein...

The major technical difficulty in gel permeation chromatography involves the careful packing of the column to ensure a uniform structure and it is absolutely essential that the level of the solvent is never allowed to fall below the level of the gel otherwise disruption of the gel bed occurs. Additionally, because most gels are compressible, only an absolute minimum of pressure should be applied to prevent alteration of the pore characteristics. In practice hydrostatic pressures greater that 30-40 cm should not be used and pumps should be used only with non-compressible gels. 

Detergents disrupt protein-lipid and protein-protein interactions and play a large role in gel electrophoresis.59 60 SDS is the most common detergent used in PAGE... 

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