Mini gel systems

An advance in the SDS-PAGE separation of proteins has been the optimization of the mini gel system. Mini gel systems are available from a number of... 

Table 1 Protein molecular weight range resolved by SDS-PAGE (using a mini gel system)...

Assemble the glass plates and cast the non-denaturing PAGE gels using the mini gel system as described in Protocol 4. 

A continuous capillary viscosity detector has been developed for use in High Performance Gel Permeation Chromatography (HPGPC). This detector has been used in conjunction with a concentration detector (DRI) to provide information on the absolute molecular weight, Mark-Houwink parameters and bulk intrinsic viscosity of polymers down to a molecular weight of about 4000. The detector was tested and used with a Waters Associates Model 150 C ALC/GPC. The combined GPC/Viscometer instrumentation was automated by means of a micro/mini-computer system which permits data acquisition/reduction for each analysis. 

SDS-polyacrylamide gel electrophoresis Protein samples were analyzed under denaturing conditions in a discontinuous gel system as described by Laemmli [13] using a 5% stacking gel and a 12% separating gel in a vertical gel system (BioRad, Mini Protean 11, CA, USA). 

The separation length is the most significant factor influencing the resolving capacity of he gel. Application of IPG IEF gel of 18 cm and second-dimension SDS-PAGE of 20 cm long allows resolution of a complex mixture with approximately 200 proteins. Rapid screening can be achieved by mini-gel formats. However, only a few hundred proteins can be separated by such system. So, the amount of the protein separated depends upon the size of the gel. 

For sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) use a Mini-Protein II slab-gel system (Bio-Rad) or equivalent. 

Stalcup aiid co-workers [14] adapted this method to a continuous elution mini-prep electrophoresis apparatus shown in Fig. 11-3. In this apparatus, the end of the electrophoretic gel is continuously washed with elution buffer. The eluent can then be monitored using an HPLC detector (Fig. 11-4) and sent to a fraction collector where the purified enantiomers, as well as the chiral additive, may be recovered. In this system, the gel configuration was approximately 100 mm x 7 mm, and was aircooled. The number of theoretical plates obtained for 0.5 mg of piperoxan with this gel was approximately 200. A larger, water-cooled gel was able to handle 15 mg of... 

Progress of the elution can be monitored by placing aliquots of the eluent on a mini-silica gel G plate, spraying with sulfuric acid and then charring. This is only one example of a solvent system that works for this particular situation. Many other combinations could be tried and again, as stated many times, it depends on the goal of the particular experiment. 

Captopril Tablets Silica gel modified with UV—Vis quaternary ammonium 2-50 pg mL-1 Flow injection system mini-column at the injection port [495]... 

Phosphorus (dissolved) Waters, sediment extracts Gel filtration UV-Vis 1.1 (low range) or 12 (high range) pgiu1 Flow injection system speciation Sephadex G-25 mini-column UV-assisted in-line sample preparation two dynamic concentration ranges [546]... 

The transfer procedure described here has been nsed with tank-based electro-transfer apparatuses such as the Bio-Rad Mini Transblot Electrophoretic Transfer Cell. Other systems will require different arrangements, buffers, and quantities, but almost all will retain the basic features of the methods described here— namely, direct contact of the gel with a nitrocellulose membrane, followed by electrophoretic transfer of proteins from the gel to the membrane. 

Capillary nonequilibrium pH gradient gel electrophoresis in conjunction with mini slab polyacrylamide electrophoresis is a rapid, cost-effective, and easy-to-handle system for the two-dimensional analysis of complex protein mixtures in one day. 

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