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Discontinuous Electrophoresis
Discontinuous electrophoresis (colloquially ''disc electrophoresis'') is a type of polyacrylamide gel electrophoresis. It was developed by Ornstein and Davis. This method produces high resolution and good band definition. It is widely used technique for separating proteins according to size and charge. Method In this method, the gel is divided into two discontinuous parts, resolving and stacking gel, both have different concentrations of polyacrylamide. The one with lower concentration is stacked on top of the one with higher concentration. Discontinuity is based on four parameters: gel structure, pH value of the buffer, ionic strength of the buffer, and the nature of the ions in the gel and electrode buffer. The electrode buffer contains glycine. Glycine has very low net charge at pH 6.8 of stacking gel, so it has low mobility. The proteins are separated according to the principle of isotachophoresis and form stacks in the order of mobility (stacking effect). Mobility depends o ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Gel Electrophoresis 2
A gel is a semi-solid that can have properties ranging from soft and weak to hard and tough. Gels are defined as a substantially dilute cross-linked system, which exhibits no flow when in the steady-state, although the liquid phase may still diffuse through this system. A gel has been defined phenomenologically as a soft, solid or solid-like material consisting of two or more components, one of which is a liquid, present in substantial quantity. By weight, gels are mostly liquid, yet they behave like solids because of a three-dimensional cross-linked network within the liquid. It is the crosslinking within the fluid that gives a gel its structure (hardness) and contributes to the adhesive stick (tack). In this way, gels are a dispersion of molecules of a liquid within a solid medium. The word ''gel'' was coined by 19th-century Scottish chemist Thomas Graham by clipping from '' gelatine''. The process of forming a gel is called gelation. IUPAC definition } ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Polyacrylamide
Polyacrylamide (abbreviated as PAM) is a polymer with the formula (-CH2CHCONH2-). It has a linear-chain structure. PAM is highly water-absorbent, forming a soft gel when hydrated. In 2008, an estimated 750,000,000 kg were produced, mainly for water treatment and the paper and mineral industries. Physicochemical properties Polyacrylamide is a polyolefin. It can be viewed as polyethylene with amide substituents on alternating carbons. Unlike various nylons, polyacrylamide is not a polyamide because the amide groups are not in the polymer backbone. Owing to the presence of the amide (CONH2) groups, alternating carbon atoms in the backbone are stereogenic (colloquially: chiral). For this reason, polyacrylamide exists in atactic, syndiotactic, and isotactic forms, although this aspect is rarely discussed. The polymerization is initiated with radicals and is assumed to be stereorandom. Copolymers and modified polymers Linear polyacrylamide is a water-soluble polymer. Other p ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Gel Electrophoresis
Gel electrophoresis is a method for separation and analysis of biomacromolecules ( DNA, RNA, proteins, etc.) and their fragments, based on their size and charge. It is used in clinical chemistry to separate proteins by charge or size (IEF agarose, essentially size independent) and in biochemistry and molecular biology to separate a mixed population of DNA and RNA fragments by length, to estimate the size of DNA and RNA fragments or to separate proteins by charge. Nucleic acid molecules are separated by applying an electric field to move the negatively charged molecules through a matrix of agarose or other substances. Shorter molecules move faster and migrate farther than longer ones because shorter molecules migrate more easily through the pores of the gel. This phenomenon is called sieving. Proteins are separated by the charge in agarose because the pores of the gel are too small to sieve proteins. Gel electrophoresis can also be used for the separation of nanoparticles. ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Glycine
Glycine (symbol Gly or G; ) is an amino acid that has a single hydrogen atom as its side chain. It is the simplest stable amino acid (carbamic acid is unstable), with the chemical formula NH2‐ CH2‐ COOH. Glycine is one of the proteinogenic amino acids. It is encoded by all the codons starting with GG (GGU, GGC, GGA, GGG). Glycine is integral to the formation of alpha-helices in secondary protein structure due to its compact form. For the same reason, it is the most abundant amino acid in collagen triple-helices. Glycine is also an inhibitory neurotransmitter – interference with its release within the spinal cord (such as during a ''Clostridium tetani'' infection) can cause spastic paralysis due to uninhibited muscle contraction. It is the only achiral proteinogenic amino acid. It can fit into hydrophilic or hydrophobic environments, due to its minimal side chain of only one hydrogen atom. History and etymology Glycine was discovered in 1820 by the French chemist He ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Electrophoretic Mobility
Electrophoresis, from Ancient Greek ἤλεκτρον (ḗlektron, "amber") and φόρησις (phórēsis, "the act of bearing"), is the motion of dispersed particles relative to a fluid under the influence of a spatially uniform electric field. Electrophoresis of positively charged particles (cations) is sometimes called cataphoresis, while electrophoresis of negatively charged particles (anions) is sometimes called anaphoresis. The electrokinetic phenomenon of electrophoresis was observed for the first time in 1807 by Russian professors Peter Ivanovich Strakhov and Ferdinand Frederic Reuss at Moscow University, who noticed that the application of a constant electric field caused clay particles dispersed in water to migrate. It is ultimately caused by the presence of a charged interface between the particle surface and the surrounding fluid. It is the basis for analytical techniques used in chemistry for separating molecules by size, charge, or binding affinity. Electropho ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Isotachophoresis
Isotachophoresis (ITP) is a technique in analytical chemistry used for selective separation and concentration of ionic analytes. It is a form of electrophoresis; charged analytes are separated based on ionic mobility, a quantity which tells how fast an ion migrates through an electric field. Overview In conventional ITP separations, a discontinuous buffer system is used. The sample is introduced between a zone of fast leading electrolyte (LE) and a zone of slow terminating (or: trailing) electrolyte (TE). Usually, the LE and the TE have a common counterion, but the co-ions (having charges with the same sign as the analytes of interest) are different: the LE is defined by co-ions with high ionic mobility, while the TE is defined by co-ions with low ionic mobility. The analytes of interest have intermediate ionic mobility. Application of an electric potential results in a low electrical field in the leading electrolyte and a high electrical field in the terminating electrolyte. ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Anode
An anode is an electrode of a polarized electrical device through which conventional current enters the device. This contrasts with a cathode, an electrode of the device through which conventional current leaves the device. A common mnemonic is ACID, for "anode current into device". The direction of conventional current (the flow of positive charges) in a circuit is opposite to the direction of electron flow, so (negatively charged) electrons flow out the anode of a galvanic cell, into an outside or external circuit connected to the cell. For example, the end of a household battery marked with a "-" (minus) is the anode. In both a galvanic cell and an electrolytic cell, the anode is the electrode at which the oxidation reaction occurs. In a galvanic cell the anode is the wire or plate having excess negative charge as a result of the oxidation reaction. In an electrolytic cell, the anode is the wire or plate upon which excess positive charge is imposed. As a result of this, anion ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Homogeneous
Homogeneity and heterogeneity are concepts often used in the sciences and statistics relating to the uniformity of a substance or organism. A material or image that is homogeneous is uniform in composition or character (i.e. color, shape, size, weight, height, distribution, texture, language, income, disease, temperature, radioactivity, architectural design, etc.); one that is heterogeneous is distinctly nonuniform in at least one of these qualities. Heterogeneous Mixtures, in chemistry, is where certain elements are unwillingly combined and, when given the option, will separate. Etymology and spelling The words ''homogeneous'' and ''heterogeneous'' come from Medieval Latin ''homogeneus'' and ''heterogeneus'', from Ancient Greek ὁμογενής (''homogenēs'') and ἑτερογενής (''heterogenēs''), from ὁμός (''homos'', “same”) and ἕτερος (''heteros'', “other, another, different”) respectively, followed by γένος (''genos'', “kind”); - ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Buffer Solution
A buffer solution (more precisely, pH buffer or hydrogen ion buffer) is an aqueous solution consisting of a mixture of a weak acid and its conjugate base, or vice versa. Its pH changes very little when a small amount of strong acid or base is added to it. Buffer solutions are used as a means of keeping pH at a nearly constant value in a wide variety of chemical applications. In nature, there are many living systems that use buffering for pH regulation. For example, the bicarbonate buffering system is used to regulate the pH of blood, and bicarbonate also acts as a buffer in the ocean. Principles of buffering Buffer solutions resist pH change because of a chemical equilibrium between the weak acid HA and its conjugate base A−: When some strong acid is added to an equilibrium mixture of the weak acid and its conjugate base, hydrogen ions (H+) are added, and the equilibrium is shifted to the left, in accordance with Le Chatelier's principle. Because of this, the hydrogen io ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Affinity Electrophoresis
Affinity electrophoresis is a general name for many analytical methods used in biochemistry and biotechnology. Both qualitative and quantitative information may be obtained through affinity electrophoresis. The methods include the so-called electrophoretic mobility shift assay, charge shift electrophoresis and affinity capillary electrophoresis. The methods are based on changes in the electrophoretic pattern of molecules (mainly macromolecules) through biospecific interaction or complex formation. The interaction or binding of a molecule, charged or uncharged, will normally change the electrophoretic properties of a molecule. Membrane proteins may be identified by a shift in mobility induced by a charged detergent. Nucleic acids or nucleic acid fragments may be characterized by their affinity to other molecules. The methods have been used for estimation of binding constants, as for instance in lectin affinity electrophoresis or characterization of molecules with specific features l ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
SDS-PAGE
SDS-PAGE (sodium dodecyl sulfate–polyacrylamide gel electrophoresis) is a discontinuous electrophoretic system developed by Ulrich K. Laemmli which is commonly used as a method to separate proteins with molecular masses between 5 and 250 kDa. The combined use of sodium dodecyl sulfate (SDS, also known as sodium lauryl sulfate) and polyacrylamide gel allows to eliminate the influence of structure and charge, and proteins are separated solely on the basis of differences in their molecular weight. Properties SDS-PAGE is an electrophoresis method that allows protein separation by mass. The medium (also referred to as ′matrix′) is a polyacrylamide-based discontinuous gel. The polyacrylamide-gel is typically sandwiched between two glass plates in a slab gel. Although tube gels (in glass cylinders) were used historically, they were rapidly made obsolete with the invention of the more convenient slab gels. In addition, SDS (sodium dodecyl sulfate) is used. About 1.4 grams of S ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Isotachophoresis
Isotachophoresis (ITP) is a technique in analytical chemistry used for selective separation and concentration of ionic analytes. It is a form of electrophoresis; charged analytes are separated based on ionic mobility, a quantity which tells how fast an ion migrates through an electric field. Overview In conventional ITP separations, a discontinuous buffer system is used. The sample is introduced between a zone of fast leading electrolyte (LE) and a zone of slow terminating (or: trailing) electrolyte (TE). Usually, the LE and the TE have a common counterion, but the co-ions (having charges with the same sign as the analytes of interest) are different: the LE is defined by co-ions with high ionic mobility, while the TE is defined by co-ions with low ionic mobility. The analytes of interest have intermediate ionic mobility. Application of an electric potential results in a low electrical field in the leading electrolyte and a high electrical field in the terminating electrolyte. ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
