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Epicocconone
Epicocconone is a long Stokes' shift fluorogenic natural product found in the fungus '' Epicoccum nigrum''. Though weakly fluorescent in water (green emission, 520 nm) it reacts covalently yet reversibly with primary amines such as those in proteins to yield a product with a strong orange-red emission (610 nm). Epicoconone is notable because it the first covalent/ reversible/turn-on fluorophore to be discovered and is a natural product with a new fluorescent scaffold. It is also cell membrane permeable, unlike many other fluorophores, and subsequently can be used in ''in vivo'' (live cell) applications. Additionally, this dye can be used as a sensitive total protein stain for 1D and 2D electrophoresis, quantitative determination of protein concentration, making it a powerful loading control for 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 ...
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Epicoccum Nigrum
''Epicoccum nigrum'' is a species of fungus in the phylum Ascomycota. A plant pathogen and endophyte, it is a widespread fungus which produces coloured pigments that can be used as antifungal agents against other pathogenic fungi. The fluorescent stain epicocconone is extracted from it. Growth and morphology ''Epicoccum nigrum'' (1825) is a fungus with no known teleomorph form. It has been classified as a member of the Hyphomycetes, in the Deuteromycota, as well as the Fungi Imperfecti because it is only known to reproduce asexually. Despite that it is not yeast-like, it has been included in the broad, unrelated category of fungi known as black yeasts. The fungus grows felty colonies in bright shades of yellow, orange, and red, often with brown or black throughout. Colonies grow quickly, reaching about 6 cm in diameter in 2 days at room temperature. Mycelia contain both chitin and cellulose. ''Epicoccum nigrum'' forms blastoconidia that are darkly coloured, warted and sphe ...
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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 antibod ...
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Fluorogenic
Fluorogenic describes a property of chemical compounds which are initially not fluorescent, but become fluorescent through a chemical reaction, typically through an intermolecular covalent reaction which binds the now fluorescent compound to a target molecule. IUPAC uses a broader definition of fluorogenic, wherein an enhancement of fluorescence via a chemical reaction is not required, however in contrast to the IUPAC definition common use of fluorogenic does not refer to non-reaction effects like the enhancement of fluorescence from a fluorophore being in different solvents. Fluorogenic labeling reagents are often used in analytical chemistry procedures, particularly in HPLC or CE to derivative target compounds (e.g. labeling the primary amines of polypeptides), thereby allowing enhanced sensitivity through fluorescence based detection. Examples * OPA * Fluorescamine Fluorescamine is a spiro compound that is not fluorescent itself, but reacts with primary amines to form highly ...
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Natural Product
A natural product is a natural compound or substance produced by a living organism—that is, found in nature. In the broadest sense, natural products include any substance produced by life. Natural products can also be prepared by chemical synthesis (both semisynthesis and total synthesis and have played a central role in the development of the field of organic chemistry by providing challenging synthetic targets). The term ''natural product'' has also been extended for commercial purposes to refer to cosmetics, dietary supplements, and foods produced from natural sources without added artificial ingredients. Within the field of organic chemistry, the definition of natural products is usually restricted to organic compounds isolated from natural sources that are produced by the pathways of primary or secondary metabolism. Within the field of medicinal chemistry, the definition is often further restricted to secondary metabolites. Secondary metabolites (or specialized meta ...
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Nanometer
330px, Different lengths as in respect to the Molecule">molecular scale. The nanometre (international spelling as used by the International Bureau of Weights and Measures; SI symbol: nm), or nanometer (American spelling Despite the various list of dialects of English, English dialects spoken from country to country and within different regions of the same country, there are only slight regional variations in English orthography, the two most notable variati ...), is a units of measurement, unit of length in the International System of Units (SI), equal to one billionth (short scale) or one thousand million (long scale) of a metre, meter (0.000000001 m) and to 1000 picometres. One nanometre can be expressed in scientific notation as 1 × 10−9 m and as  m. History The nanometre was formerly known as the "''millimicrometre''" – or, more commonly, the "''millimicron''" for short – since it is of a micrometre, micrometer. It was often de ...
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Covalent Bond
A covalent bond is a chemical bond that involves the sharing of electrons to form electron pairs between atoms. These electron pairs are known as shared pairs or bonding pairs. The stable balance of attractive and repulsive forces between atoms, when they share electrons, is known as covalent bonding. For many molecules, the sharing of electrons allows each atom to attain the equivalent of a full valence shell, corresponding to a stable electronic configuration. In organic chemistry, covalent bonding is much more common than ionic bonding. Covalent bonding also includes many kinds of interactions, including σ-bonding, π-bonding, metal-to-metal bonding, agostic interactions, bent bonds, three-center two-electron bonds and three-center four-electron bonds. The term "covalence" was introduced by Irving Langmuir in 1919, with Nevil Sidgwick using "co-valent link" in the 1920s. Merriam-Webster dates the specific phrase ''covalent bond'' to 1939, recognizing its first known ...
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Reversible Reaction
A reversible reaction is a reaction in which the conversion of reactants to products and the conversion of products to reactants occur simultaneously. : \mathit aA + \mathit bB \mathit cC + \mathit dD A and B can react to form C and D or, in the reverse reaction, C and D can react to form A and B. This is distinct from a reversible process in thermodynamics. Weak acids and bases undergo reversible reactions. For example, carbonic acid: : H2CO3 (l) + H2O(l) ⇌ HCO3−(aq) + H3O+(aq). The concentrations of reactants and products in an equilibrium mixture are determined by the analytical concentrations of the reagents (A and B or C and D) and the equilibrium constant, ''K''. The magnitude of the equilibrium constant depends on the Gibbs free energy change for the reaction. So, when the free energy change is large (more than about 30 kJ mol−1), the equilibrium constant is large (log K > 3) and the concentrations of the reactants at equilibrium are very small. Such a reaction i ...
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Primary Amines
In chemistry, amines (, ) are organic compounds that contain carbon-nitrogen bonds. Amines are formed when one or more hydrogen atoms in ammonia are replaced by alkyl or aryl groups. The nitrogen atom in an amine possesses a lone pair of electrons. Amines can also exist as hetero cyclic compounds. Aniline is the simplest aromatic amine, consisting of a benzene ring bonded to an amino group. Amines are classified into three types: primary (1°), secondary (2°), and tertiary (3°) amines. Primary amines (1°) contain one alkyl or aryl substituent and have the general formula RNH2. Secondary amines (2°) have two alkyl or aryl groups attached to the nitrogen atom, with the general formula R2NH. Tertiary amines (3°) contain three substituent groups bonded to the nitrogen atom, and are represented by the formula R3N. The functional group present in primary amines is called the amino group. Classification of amines Amines can be classified according to the nature and number o ...
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Protein
Proteins are large biomolecules and macromolecules that comprise one or more long chains of amino acid residue (biochemistry), residues. Proteins perform a vast array of functions within organisms, including Enzyme catalysis, catalysing metabolic reactions, DNA replication, Cell signaling, responding to stimuli, providing Cytoskeleton, structure to cells and Fibrous protein, organisms, and Intracellular transport, transporting molecules from one location to another. Proteins differ from one another primarily in their sequence of amino acids, which is dictated by the Nucleic acid sequence, nucleotide sequence of their genes, and which usually results in protein folding into a specific Protein structure, 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 pep ...
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Orange (colour)
Orange is the colour between yellow and red on the spectrum of visible light. The human eyes perceive orange when observing light with a dominant wavelength between roughly 585 and 620 nanometres. In traditional colour theory, it is a secondary colour of pigments, produced by mixing yellow and red. In the RGB colour model, it is a tertiary colour. It is named after the orange (fruit), fruit of the same name. The orange colour of many fruits and vegetables, such as carrots, pumpkins, sweet potatoes, and Orange (fruit), oranges, comes from carotenes, a type of photosynthetic pigment. These pigments convert the light energy that the plants absorb from the Sun into chemical energy for the plants' growth. Similarly, the hues of autumn leaves are from the same pigment after chlorophyll is removed. In Europe and the United States, surveys show that orange is the colour most associated with amusement, the unconventional, extroversion, warmth, fire, energy, activity, danger, taste and a ...
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