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Dot Blot
A dot blot (or slot blot) is a technique in molecular biology used to detect proteins. It represents a simplification of the western blot method, with the exception that the proteins to be detected are not first separated by electrophoresis. Instead, the sample is applied directly on a membrane in a single spot, and the blotting procedure is performed. The technique offers significant savings in time, as chromatography or gel electrophoresis, and the complex blotting procedures for the gel are not required. However, it offers no information on the size of the target protein. Uses Performing a dot blot is similar in idea to performing a western blot, with the advantage of faster speed and lower cost. Dot blots are also performed to screen the binding capabilities of an antibody. Methodology A general dot blot protocol involves spotting 1–2 microliters of a samples onto a nitrocellulose Nitrocellulose (also known as cellulose nitrate, flash paper, flash cotton, guncot ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Dot Blot De ADN
A dot is usually a small, round spot. Dot, DoT or DOT may also refer to: Orthography * Full stop or "period", a sentence terminator * Dot (diacritic), a mark above or below a character (e.g. ȧ, ạ, İ, Ċ, ċ, etc.), usually to indicate sound mutation * Interpunct () also known as an "interpoint", "middle dot", "middot", "centered dot" or "centred dot", a punctuation mark Mathematics * "Dot as decimal point", the form of decimal separator generally used in anglophone countries, e.g., $24.99 * Dot operator, a notation used to signify multiplication * Dot product, algebraic operation returning a single number from two equal-length sequences Arts and entertainment * ''The Dot'' (Toyism), a work of art in Emmen, the Netherlands * "Dot" (song), a single by the Los Angeles pop punk band ALL * "Dot", a song by Destiny's Child featured on the ''Charlie's Angels'' soundtrack * '' Dot, A Drama in Three Acts'', an 1859 stage adaption of the book ''The Cricket on the Hearth'' by Ch ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Molecular Biology
Molecular biology is a branch of biology that seeks to understand the molecule, molecular basis of biological activity in and between Cell (biology), cells, including biomolecule, biomolecular synthesis, modification, mechanisms, and interactions. Though cells and other microscopic structures had been observed in living organisms as early as the 18th century, a detailed understanding of the mechanisms and interactions governing their behavior did not emerge until the 20th century, when technologies used in physics and chemistry had advanced sufficiently to permit their application in the biological sciences. The term 'molecular biology' was first used in 1945 by the English physicist William Astbury, who described it as an approach focused on discerning the underpinnings of biological phenomena—i.e. uncovering the physical and chemical structures and properties of biological molecules, as well as their interactions with other molecules and how these interactions explain observ ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
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 ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Electrophoresis
Electrophoresis is the motion of charged dispersed particles or dissolved charged molecules relative to a fluid under the influence of a spatially uniform electric field. As a rule, these are zwitterions with a positive or negative net charge. Electrophoresis is used in laboratories to separate macromolecules based on their charges. The technique normally applies a negative charge called cathode so anionic protein molecules move towards a positive charge called anode. Therefore, electrophoresis of positively charged particles or molecules (cations) is sometimes called cataphoresis, while electrophoresis of negatively charged particles or molecules (anions) is sometimes called anaphoresis. Electrophoresis is the basis for analytical techniques used in biochemistry and molecular biology to separate particles, molecules, or ions by size, charge, or binding affinity, either freely or through a supportive medium using a one-directional flow of electrical charge. It is use ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Chromatography
In chemical analysis, chromatography is a laboratory technique for the Separation process, separation of a mixture into its components. The mixture is dissolved in a fluid solvent (gas or liquid) called the ''mobile phase'', which carries it through a system (a column, a capillary tube, a plate, or a sheet) on which a material called the ''stationary phase'' is fixed. Because the different constituents of the mixture tend to have different affinities for the stationary phase and are retained for different lengths of time depending on their interactions with its surface sites, the constituents travel at different apparent velocities in the mobile fluid, causing them to separate. The separation is based on the differential partitioning between the mobile and the stationary phases. Subtle differences in a compound's partition coefficient result in differential retention on the stationary phase and thus affect the separation. Chromatography may be ''preparative'' or ''analytical' ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Gel Electrophoresis
Gel electrophoresis is an electrophoresis method for separation and analysis of biomacromolecules (DNA, RNA, proteins, etc.) and their fragments, based on their size and charge through a gel. 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 gel matrix of agarose, polyacrylamide, 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 large to sieve proteins. Gel electrophoresi ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Nitrocellulose
Nitrocellulose (also known as cellulose nitrate, flash paper, flash cotton, guncotton, pyroxylin and flash string, depending on form) is a highly flammable compound formed by nitrating cellulose through exposure to a mixture of nitric acid and sulfuric acid. One of its first major uses was as guncotton, a replacement for gunpowder as propellant in firearms. It was also used to replace gunpowder as a low-order explosive in mining and other applications. In the form of collodion, it was also a critical component in an early photographic emulsion, the use of which revolutionized photography in the 1860s. In the 20th century, it was adapted to automobile lacquer and adhesives. Production The process uses a mixture of nitric acid and sulfuric acid to convert cellulose into nitrocellulose. The quality of the cellulose is important. Hemicellulose, lignin, pentosans, and mineral salts give inferior nitrocelluloses. In organic chemistry, nitrocellulose is a nitrate ester, not a ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
PVDF
Polyvinylidene fluoride or polyvinylidene difluoride (PVDF) is a highly non-reactive thermoplastic fluoropolymer produced by the polymerization of vinylidene difluoride. Its chemical formula is (C2H2F2)''n''. PVDF is a specialty plastic used in applications requiring the highest purity, as well as resistance to solvents, acids and hydrocarbons. PVDF has low density 1.78 g/cm3 in comparison to other fluoropolymers, like polytetrafluoroethylene. It is available in the form of piping products, sheet, tubing, films, plate and an insulator for premium wire. It can be injected, molded or welded and is commonly used in the chemical, semiconductor, medical and defense industries, as well as in lithium-ion batteries. It is also available as a cross-linked closed-cell foam, used increasingly in aviation and aerospace applications, and as an exotic 3D printer filament. It can also be used in repeated contact with food products, as it is FDA-compliant and non-toxic below its degr ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Chemiluminescent
Chemiluminescence (also chemoluminescence) is the emission of light (luminescence) as the result of a chemical reaction, i.e. a chemical reaction results in a flash or glow of light. A standard example of chemiluminescence in the laboratory setting is the luminol test. Here, blood is indicated by luminescence upon contact with iron in hemoglobin. When chemiluminescence takes place in living organisms, the phenomenon is called bioluminescence. A light stick emits light by chemiluminescence. Physical description As in many chemical reactions, chemiluminescence starts with the combining of two compounds, say A and B, to give a product C. Unlike most chemical reactions, the product C converts to a further product, which is produced in an electronically excited state often indicated with an asterisk: : A + B → C : C → D* D* then emits a photon (''h''ν), to give the ground state of D: I : D* → D + ''h''ν In theory, one photon of light should be given off for each mo ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
ELISpot
The enzyme-linked immunosorbent spot (ELISpot) is a type of assay that focuses on quantitatively measuring the frequency of cytokine secretion for a single cell. The ELISpot Assay is also a form of immunostaining since it is classified as a technique that uses antibodies to detect a protein analyte, with the word analyte referring to any biological or chemical substance being identified or measured. The FluoroSpot Assay is a variation of the ELISpot assay. The FluoroSpot Assay uses fluorescence in order to analyze multiple analytes, meaning it can detect the secretion of more than one type of protein. History Cecil Czerkinsky first described ELISpot in 1983 as a new way to quantify the production of an antigen-specific immunoglobulin by hybridoma cells. In 1988, Czerkinsky developed an ELISA spot assay that quantified the secretion of a lymphokine by T cells. In the same year, dual-color ELISpot was combined with computer imaging for the first time, which allowed for the enum ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
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 ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Genetics Techniques
Genetics is the study of genes, genetic variation, and heredity in organisms.Hartl D, Jones E (2005) It is an important branch in biology because heredity is vital to organisms' evolution. Gregor Mendel, a Moravian Augustinian friar working in the 19th century in Brno, was the first to study genetics scientifically. Mendel studied "trait inheritance", patterns in the way traits are handed down from parents to offspring over time. He observed that organisms (pea plants) inherit traits by way of discrete "units of inheritance". This term, still used today, is a somewhat ambiguous definition of what is referred to as a gene. Trait inheritance and molecular inheritance mechanisms of genes are still primary principles of genetics in the 21st century, but modern genetics has expanded to study the function and behavior of genes. Gene structure and function, variation, and distribution are studied within the context of the cell, the organism (e.g. dominance), and within the ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
