Protein Electrophoresis
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Protein Electrophoresis
Protein electrophoresis is a method for analysing the proteins in a fluid or an extract. The electrophoresis may be performed with a small volume of sample in a number of alternative ways with or without a supporting medium: SDS polyacrylamide gel electrophoresis (in short: gel electrophoresis, PAGE, or SDS-electrophoresis), free-flow electrophoresis, electrofocusing, isotachophoresis, affinity electrophoresis, immunoelectrophoresis, counterelectrophoresis, and capillary electrophoresis. Each method has many variations with individual advantages and limitations. Gel electrophoresis is often performed in combination with electroblotting immunoblotting to give additional information about a specific protein. Because of practical limitations, protein electrophoresis is generally not suited as a preparative method. Denaturing gel methods SDS-PAGE SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis, describes a collection of related techniques to separate proteins a ...
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Detergent
A detergent is a surfactant or a mixture of surfactants with cleansing properties when in dilute solutions. There are a large variety of detergents, a common family being the alkylbenzene sulfonates, which are soap-like compounds that are more soluble in hard water, because the polar sulfonate (of detergents) is less likely than the polar carboxylate (of soap) to bind to calcium and other ions found in hard water. Definitions The word ''detergent'' is derived from the Latin adjective ''detergens'', from the verb ''detergere'', meaning to wipe or polish off. Detergent is a surfactant or a mixture of surfactants with cleansing properties when in dilute solutions. However, conventionally, detergent is used to mean synthetic cleaning compounds as opposed to ''soap'' (a salt of the natural fatty acid), even though soap is also a detergent in the true sense. In domestic contexts, the term ''detergent'' refers to household cleaning products such as ''laundry detergent'' or '' dish ...
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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 residues ...
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Chlorophyll
Chlorophyll (also chlorophyl) is any of several related green pigments found in cyanobacteria and in the chloroplasts of algae and plants. Its name is derived from the Greek words , ("pale green") and , ("leaf"). Chlorophyll allow plants to absorb energy from light. Chlorophylls absorb light most strongly in the blue portion of the electromagnetic spectrum as well as the red portion. Conversely, it is a poor absorber of green and near-green portions of the spectrum. Hence chlorophyll-containing tissues appear green because green light, diffusively reflected by structures like cell walls, is less absorbed. Two types of chlorophyll exist in the photosystems of green plants: chlorophyll ''a'' and ''b''. History Chlorophyll was first isolated and named by Joseph Bienaimé Caventou and Pierre Joseph Pelletier in 1817. The presence of magnesium in chlorophyll was discovered in 1906, and was that element's first detection in living tissue. After initial work done by German chemi ...
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Heme
Heme, or haem (pronounced / hi:m/ ), is a precursor to hemoglobin, which is necessary to bind oxygen in the bloodstream. Heme is biosynthesized in both the bone marrow and the liver. In biochemical terms, heme is a coordination complex "consisting of an iron ion coordinated to a porphyrin acting as a tetradentate ligand, and to one or two axial ligands." The definition is loose, and many depictions omit the axial ligands. Among the metalloporphyrins deployed by metalloproteins as prosthetic groups, heme is one of the most widely used and defines a family of proteins known as hemoproteins. Hemes are most commonly recognized as components of hemoglobin, the red pigment in blood, but are also found in a number of other biologically important hemoproteins such as myoglobin, cytochromes, catalases, heme peroxidase, and endothelial nitric oxide synthase. The word ''haem'' is derived from Greek ''haima'' meaning "blood". Function Hemoproteins have diverse biological functions incl ...
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Prosthetic Group
A prosthetic group is the non-amino acid component that is part of the structure of the heteroproteins or conjugated proteins, being tightly linked to the apoprotein. Not to be confused with the cofactor that binds to the enzyme apoenzyme (either a holoprotein or heteroprotein) by non-covalent binding a non-protein (non-amino acid) This is a component of a conjugated protein that is required for the protein's biological activity. The prosthetic group may be organic (such as a vitamin, sugar, RNA, phosphate or lipid) or inorganic (such as a metal ion). Prosthetic groups are bound tightly to proteins and may even be attached through a covalent bond. They often play an important role in enzyme catalysis. A protein without its prosthetic group is called an apoprotein, while a protein combined with its prosthetic group is called a holoprotein. A non-covalently bound prosthetic group cannot generally be removed from the holoprotein without denaturating the protein. Thus, the term ...
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Fluorescence
Fluorescence is the emission of light by a substance that has absorbed light or other electromagnetic radiation. It is a form of luminescence. In most cases, the emitted light has a longer wavelength, and therefore a lower photon energy, than the absorbed radiation. A perceptible example of fluorescence occurs when the absorbed radiation is in the ultraviolet region of the electromagnetic spectrum (invisible to the human eye), while the emitted light is in the visible region; this gives the fluorescent substance a distinct color that can only be seen when the substance has been exposed to UV light. Fluorescent materials cease to glow nearly immediately when the radiation source stops, unlike phosphorescent materials, which continue to emit light for some time after. Fluorescence has many practical applications, including mineralogy, gemology, medicine, chemical sensors (fluorescence spectroscopy), fluorescent labelling, dyes, biological detectors, cosmic-ray detection, vacu ...
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Western Blot
The western blot (sometimes called the protein immunoblot), or western blotting, is a widely used analytical technique in molecular biology and immunogenetics to detect specific proteins in a sample of tissue homogenate or extract. Besides detecting the proteins, this technique is also utilized to visualize, distinguish, and quantify the different proteins in a complicated protein combination. Western blot technique uses three elements to achieve its task of separating a specific protein from a complex: separation by size, transfer of protein to a solid support, and marking target protein using a primary and secondary antibody to visualize. A synthetic or animal-derived antibody (known as the primary antibody) is created that recognizes and binds to a specific target protein. The electrophoresis membrane is washed in a solution containing the primary antibody, before excess antibody is washed off. A secondary antibody is added which recognizes and binds to the primary antibody ...
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Chemoluminescence
Chemiluminescence (also chemoluminescence) is the emission of light (luminescence) as the result of a chemical reaction. There may also be limited emission of heat. Given reactants A and B, with an excited intermediate ◊, : + -> lozenge -> roducts+ light For example, if is luminol and is hydrogen peroxide in the presence of a suitable catalyst we have: :\underset + \underset -> 3-APAlozenge-> + light where: * 3-APA is 3-aminophthalate * 3-APA ''◊is the vibronic excited state fluorescing as it decays to a lower energy level. General description The decay of this excited state ''◊to a lower energy level causes light emission. In theory, one photon of light should be given off for each molecule of reactant. This is equivalent to the Avogadro number of photons per mole of reactant. In actual practice, non-enzymatic reactions seldom exceed 1% QC, quantum efficiency. In a chemical reaction, reactants collide to form a transition state, the enthalpic maximum in ...
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Quenching (fluorescence)
Quenching refers to any process which decreases the fluorescence intensity of a given substance. A variety of processes can result in quenching, such as excited state reactions, energy transfer, complex-formation and collisional quenching. As a consequence, quenching is often heavily dependent on pressure and temperature. Molecular oxygen, iodide ions and acrylamide are common chemical quenchers. The chloride ion is a well known quencher for quinine fluorescence. Quenching poses a problem for non-instant spectroscopic methods, such as laser-induced fluorescence. Quenching is made use of in optode sensors; for instance the quenching effect of oxygen on certain ruthenium complexes allows the measurement of oxygen saturation in solution. Quenching is the basis for Förster resonance energy transfer (FRET) assays. Quenching and dequenching upon interaction with a specific molecular biological target is the basis for activatable optical contrast agents for molecular imaging. Many dyes ...
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Dissociate
Dissociation in chemistry is a general process in which molecules (or ionic compounds such as salts, or complexes) separate or split into other things such as atoms, ions, or radicals, usually in a reversible manner. For instance, when an acid dissolves in water, a covalent bond between an electronegative atom and a hydrogen atom is broken by heterolytic fission, which gives a proton (H+) and a negative ion. Dissociation is the opposite of association or recombination. Dissociation constant For reversible dissociations in a chemical equilibrium :AB A + B the dissociation constant ''K''d is the ratio of dissociated to undissociated compound :K_d = \mathrm where the brackets denote the equilibrium concentrations of the species. Dissociation degree The dissociation degree \alpha is the fraction of original solute molecules that have dissociated. It is usually indicated by the Greek symbol α. More accurately, degree of dissociation refers to the amount of solute dissociated int ...
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Charge (physics)
In physics, a charge is any of many different quantities, such as the electric charge in electromagnetism or the color charge in quantum chromodynamics. Charges correspond to the time-invariant generators of a symmetry group, and specifically, to the generators that commute with the Hamiltonian. Charges are often denoted by the letter ''Q'', and so the invariance of the charge corresponds to the vanishing commutator ,H0, where H is the Hamiltonian. Thus, charges are associated with conserved quantum numbers; these are the eigenvalues ''q'' of the generator ''Q''. Abstract definition Abstractly, a charge is any generator of a continuous symmetry of the physical system under study. When a physical system has a symmetry of some sort, Noether's theorem implies the existence of a conserved current. The thing that "flows" in the current is the "charge", the charge is the generator of the (local) symmetry group. This charge is sometimes called the Noether charge. Thus, for exampl ...
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