Epicocconone
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Epicocconone
Epicocconone is a long Stokes shift, Stokes' shift Fluorophore, fluorescent dye found in the fungus ''Epicoccum nigrum''. Though weakly fluorescent in water (green emission, 520 nanometer, nm) it reacts Reversible reaction, reversibly with proteins to yield a product with a strong Orange (colour), orange-red emission (610 nm). Epicoconone is notable because it the first covalent/Reversible reaction, reversible/turn-on fluorophore to be discovered and is a natural product with a new fluorescent scaffold. Accordingly, this dye can be used as a sensitive total protein stain for 1D and Two-dimensional gel electrophoresis, 2D electrophoresis, Quantitative analysis (chemistry), quantitative determination of protein concentration, making it a powerful loading control for Western blots. Synthetic variant In addition to the natural variant from the fungus, there are several synthetic analogs. With respect to protein staining properties there are few differences between natural and ...
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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 Fungicide, antifungal agents against other pathogenic fungi. The Fluorophore, 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 Blastoconidium, blastoconidia tha ...
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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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Stokes Shift
__NOTOC__ Stokes shift is the difference (in energy, wavenumber or frequency units) between positions of the band maxima of the absorption and emission spectra (fluorescence and Raman being two examples) of the same electronic transition. It is named after Irish physicist George Gabriel Stokes. Sometimes Stokes shifts are given in wavelength units, but this is less meaningful than energy, wavenumber or frequency units because it depends on the absorption wavelength. For instance, a 50 nm Stokes shift from absorption at 300 nm is larger in terms of energy than a 50 nm Stokes shift from absorption at 600 nm. When a system (be it a molecule or atom) absorbs a photon, it gains energy and enters an excited state. One way for the system to relax is to emit a photon, thus losing its energy (another method would be the loss of energy as translational mode energy (via vibrational-translational or electronic-translational collisional processes with other atoms or molecules)). When the ...
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Fluorophore
A fluorophore (or fluorochrome, similarly to a chromophore) is a fluorescent chemical compound that can re-emit light upon light excitation. Fluorophores typically contain several combined aromatic groups, or planar or cyclic molecules with several π bonds. Fluorophores are sometimes used alone, as a tracer in fluids, as a dye for staining of certain structures, as a substrate of enzymes, or as a probe or indicator (when its fluorescence is affected by environmental aspects such as polarity or ions). More generally they are covalently bonded to a macromolecule, serving as a marker (or dye, or tag, or reporter) for affine or bioactive reagents (antibodies, peptides, nucleic acids). Fluorophores are notably used to stain tissues, cells, or materials in a variety of analytical methods, i.e., fluorescent imaging and spectroscopy. Fluorescein, via its amine-reactive isothiocyanate derivative fluorescein isothiocyanate (FITC), has been one of the most popular fluorophores. Fro ...
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Nanometer
330px, Different lengths as in respect to the molecular scale. The nanometre (international spelling as used by the International Bureau of Weights and Measures; SI symbol: nm) or nanometer (American and British English spelling differences#-re, -er, American spelling) is a units of measurement, unit of length in the International System of Units (SI), equal to one billionth (short scale) of a metre () and to 1000 picometres. One nanometre can be expressed in scientific notation as , and as  metres. History The nanometre was formerly known as the millimicrometre – or, more commonly, the millimicron for short – since it is of a micron (micrometre), and was often denoted by the symbol mμ or (more rarely and confusingly, since it logically should refer to a ''millionth'' of a micron) as μμ. Etymology The name combines the SI prefix ''nano-'' (from the Ancient Greek , ', "dwarf") with the parent unit name ''metre'' (from Greek , ', "unit of measurement"). ...
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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 reac ...
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Protein
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 residue ...
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Orange (colour)
Orange is the colour between yellow and red on the visible spectrum, spectrum of light, visible light. 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 America, surveys show that orange is the colour most associated with amusement, the unconventional, extroversion, warmth, fire, energy, activity, danger ...
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Two-dimensional Gel Electrophoresis
Two-dimensional gel electrophoresis, abbreviated as 2-DE or 2-D electrophoresis, is a form of gel electrophoresis commonly used to analyze proteins. Mixtures of proteins are separated by two properties in two dimensions on 2D gels. 2-DE was first independently introduced by O'Farrell and Klose in 1975. Basis for separation 2-D electrophoresis begins with electrophoresis in the first dimension and then separates the molecules perpendicularly from the first to create an electropherogram in the second dimension. In electrophoresis in the first dimension, molecules are separated linearly according to their isoelectric point. In the second dimension, the molecules are then separated at 90 degrees from the first electropherogram according to molecular mass. Since it is unlikely that two molecules will be similar in two distinct properties, molecules are more effectively separated in 2-D electrophoresis than in 1-D electrophoresis. The two dimensions that proteins are separated into ...
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Quantitative Analysis (chemistry)
In analytical chemistry, quantitative analysis is the determination of the absolute or relative abundance (often expressed as a concentration) of one, several or all particular substance(s) present in a sample. Methods Once the presence of certain substances in a sample is known, the study of their absolute or relative abundance could help in determining specific properties. Knowing the composition of a sample is very important, and several ways have been developed to make it possible, like gravimetric and volumetric analysis. Gravimetric analysis yields more accurate data about the composition of a sample than volumetric analysis but also takes more time to perform in the laboratory. Volumetric analysis, on the other hand, doesn't take that much time and can produce satisfactory results. Volumetric analysis can be simply a titration based in a neutralization reaction but it can also be a precipitation or a complex forming reaction as well as a titration based in a redox reactio ...
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