AIMS Bile is an important body fluid which assists in the digestion of fat and excretion of endogenous and exogenous compounds. In the present study, an improved sample preparation for human bile was established. METHODS AND MATERIAL The method involved acetone precipitation followed by protein extraction using commercially available 2D Clean-Up kit. The effectiveness was evaluated by 2-dimen...

Electrophoresis is the main metod for separating large polyelectrolyte molecules, primarily used on DNA, in biochemistry and medicine research. An overwiev of the models of physical mechanisms that are behind this phenomena will be presented. The prominent role is taken by reptation mechanisms, by which large ‡exible polyelectrolytes thread their way through the pores of the gel matrix. There a...

The separation of polypeptides by electrophoresis allowed Arne Tiselius to describe protein fractions corresponding to albumin, -, -, and -globulins in serum with the Rrst published diagram of human serum protein electrophoresis in 1939. The number of fractions slowly expanded into electrophoretic subfractions identiRed as 1, 2, 1, 2, 1 and 2. The mobility characteristics of these fractions are...

The discovery that antigen}antibody interaction could be produced not only in liquids, but also on gel media, such as agar or agarose gels, with the formation of insoluble immunoprecipitates, opened the door to the development of the gel diffusion techniques for immunoprecipitation analysis. The Rrst of these techniques, known as double diffusion, was introduced by Ouchterlony in 1948. In this ...

INTRODUCTIONThis protocol describes a method for concentrating gel bands or spots excised from 1D or 2D acrylamide gels. Several replicate protein bands (or spots) are often required to provide sufficient material for sequencing. In these situations, it is necessary to excise several stained protein bands or spots from the gels and pre-concentrate them prior to electroblotting for amino-termina...

In the present study, we compared six different solubilization buffers and optimized two-dimensional electrophoresis (2-DE) conditions for human lymph node proteins. In addition, we developed a simple protocol for 2-D gel storage. Efficient solubilization was obtained with lysis buffers containing (a) 8 M urea, 4% CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate), 40 mM Tris ba...

Two-dimensional (2D) polyacrylamide gel electrophoresis is a classical technique for the separation of proteins. It Rrst appeared in the mid-1970s but for a long time it only found limited applications. Recently it has enjoyed an impressive renaissance. The major reasons for this are the introduction of the immobilized pH gradient (IPG) strips and the development of analytical methods capable o...

results the results showed that nl1-gc/ls2 primer set could yield species-specific amplicons, which were well distinguished and allowed better species discrimination than that generated by the its3-gc/its4 primer set, in both dgge and ttge profiles. all five candida species were discriminated by dgge and ttge using the nl1-gc/ls2 primer set. conclusions comparison of dgge and ttge profiles obta...

The most common technique for sorting, identifying, and purifying nucleic acid fragments is agarose gel electrophoresis. performance of the electrophoresis could be impacted by buffers’ composition, ionic strength, pH characteristics [1].