PAGE Is the Most Widely Used Technique for the Separation and Biochemical Analysis of Biomolecules

Description:
Tight regulation of pH is critical for the structure and function of cells and organelles. The pH environment changes dramatically along the endocytic pathway, an internalization transport process that is ‘hijacked’ by many intracellular active bacterial exotoxins, including the anthrax toxin. Here, we investigate the role of pH on single-channel properties of the anthrax toxin protective antigen (PA63). Using conductance and current noise analysis, blocker binding, ion selectivity, and poly(ethylene glycol) partitioning measurements, we show that the channel exists in two different open states (‘maximum’ and ‘main’) at pH ≥ 5.5, while only a maximum conductance state is detected at pH < 5.5. We describe two substantially distinct patterns of PA63 conductance dependence on KCl concentration uncovered at pH 6.5 and 4.5. Microscopic imaging has revolutionized our understanding of the structure and function of cells and tissues. At its best, microscopy can detect cellular components with high sensitivity and spatial resolution. Additionally, live imaging allows dynamic cellular processes to be studied in a non-invasive manner, with high temporal resolution. In the last decade, a small number of chemical reactions have been developed which occur with high efficiency and specificity under mild conditions, in aqueous media. These reactions are bio-orthogonal, involving chemical groups not found in cells and inert toward naturally occurring chemical groups. These reactions are thus ideal for labeling and detecting molecules in vivo.
One focus of our lab is to develop chemical technologies for metabolic labeling and direct imaging of biological molecules, based on these bio-orthogonal chemical reactions. My talk will present our recent work on devising novel biosynthetic probes to image nucleic acids and lipids in cells and in tissues. PAGE is the most widely used technique for the separation and biochemical analysis of biomolecules. The ever growing field of proteomics and genomics necessitates the analysis of many proteins and nucleic acid samples to understand further about the structure and function of cells. Simultaneous analysis of multiple protein samples often requires casting of many PAGE gels. Several variants of multi-gel casting/electrophoresis apparatuses are frequently used in research laboratories. Requirement of supplementary gels to match the growing demand for analysing additional protein samples sometimes become a cause of concern. Available apparatuses are not amenable to and therefore, not recommended for any modification to accommodate additional gel casting units other than what is prescribed by the manufacturer. A novel apparatus is described here for casting multiple PAGE gels comprising four detachable components that provide enhanced practicability and performance of the apparatus. This newly modified apparatus promises to be a reliable source for making multiple gels in less time without hassle. Synchronized functioning of unique components broaden the possibilities of developing inexpensive, safe, and time-saving multi-gel casting apparatus. This apparatus can be easily fabricated and modified to accommodate desired number of gel casting units. The estimated cost (∼$300) for fabrication of the main apparatus is very competitive and effortless assembly procedure can be completed within ∼30 min.
With Regards
Enderson
Journal Coordinator
Global Journal of Research and Review