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Chromatofocusing
Chromatofocusing is a protein-separation technique that allows resolution of single proteins and other ampholytes from a complex mixture according to differences in their isoelectric point The isoelectric point (pI, pH(I), IEP), is the pH at which a molecule carries no net electrical charge or is electrically neutral in the statistical mean. The standard nomenclature to represent the isoelectric point is pH(I). However, pI is also .... Chromatofocusing utilizes ion exchange resins and is typically performed on fast protein liquid chromatography (FPLC) or similar equipment capable of producing continuous buffer gradients though this is not a requirement. In contrast to typical ion exchange chromatography, where bound molecules are eluted from the resin by increasing the ionic strength of the buffer environment, chromatofocusing elutes bound species by altering the pH of the buffer. This changes the net surface charge of bound molecules, altering their avidity for the resin. As the ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Proteins
Proteins are large biomolecules and macromolecules that comprise one or more long chains of amino acid residues. Proteins perform a vast array of functions within organisms, including catalysing metabolic reactions, DNA replication, responding to stimuli, providing structure to cells and organisms, and transporting molecules from one location to another. Proteins differ from one another primarily in their sequence of amino acids, which is dictated by the nucleotide sequence of their genes, and which usually results in protein folding into a specific 3D structure that determines its activity. A linear chain of amino acid residues is called a polypeptide. A protein contains at least one long polypeptide. Short polypeptides, containing less than 20–30 residues, are rarely considered to be proteins and are commonly called peptides. The individual amino acid residues are bonded together by peptide bonds and adjacent amino acid residues. The sequence of amino acid residu ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Ampholytes
In chemistry, an amphoteric compound () is a molecule or ion that can react both as an acid and as a base. What exactly this can mean depends on which definitions of acids and bases are being used. One type of amphoteric species are amphiprotic molecules, which can either donate or accept a proton (). This is what "amphoteric" means in Brønsted–Lowry acid–base theory. Examples include amino acids and proteins, which have amine () and carboxylic acid () groups, and self-ionizable compounds such as water. Ampholytes are amphoteric molecules that contain both acidic and basic functional groups. For example, an amino acid has both a basic group and an acidic group , and exists as several structures in chemical equilibrium: :H2N-RCH-CO2H + H2O H2N-RCH-COO- + H3O+ H3N+-RCH-COOH + OH- H3N+-RCH-COO- + H2O In approximately neutral aqueous solution (pH ≅ 7), the basic amino group is mostly protonated and the carboxylic acid is mostly deprotonated, so that the predominant ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Isoelectric Point
The isoelectric point (pI, pH(I), IEP), is the pH at which a molecule carries no net electrical charge or is electrically neutral in the statistical mean. The standard nomenclature to represent the isoelectric point is pH(I). However, pI is also used. For brevity, this article uses pI. The net charge on the molecule is affected by pH of its surrounding environment and can become more positively or negatively charged due to the gain or loss, respectively, of protons (H+). Surfaces naturally charge to form a double layer. In the common case when the surface charge-determining ions are H+/HO−, the net surface charge is affected by the pH of the liquid in which the solid is submerged. The pI value can affect the solubility of a molecule at a given pH. Such molecules have minimum solubility in water or salt solutions at the pH that corresponds to their pI and often precipitate out of solution. Biological amphoteric molecules such as proteins contain both acidic and basic funct ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
