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Native PAGE
Polyacrylamide gel electrophoresis (PAGE) is a technique widely used in biochemistry, forensic chemistry, genetics, molecular biology and biotechnology to separate biological macromolecules, usually proteins or nucleic acids, according to their Electrophoresis, electrophoretic mobility. Electrophoretic mobility is a function of the length, conformation, and charge of the molecule. Polyacrylamide gel electrophoresis is a powerful tool used to analyze RNA samples. When polyacrylamide gel is denatured after electrophoresis, it provides information on the sample composition of the RNA species. Hydration reaction, Hydration of acrylonitrile results in formation of acrylamide molecules () by nitrile hydratase. Acrylamide monomer is in a powder state before addition of water. Acrylamide is toxic to the human nervous system, therefore all safety measures must be followed when working with it. Acrylamide is soluble in water and upon addition of free-radical initiators it polymerizes resu ...
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SDS-PAGE
SDS-PAGE (sodium dodecyl sulfate–polyacrylamide gel electrophoresis) is a Discontinuous electrophoresis, 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 Kilodalton, kDa. The combined use of sodium dodecyl sulfate (SDS, also known as sodium lauryl sulfate) and polyacrylamide gel eliminates the influence of structure and charge, and proteins are separated by differences in their size. At least up to 2012, the publication describing it was the most frequently cited paper by a single author, and the second most cited overall. 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 rapid ...
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Native State
In biochemistry, the native state of a protein or nucleic acid is its properly Protein folding, folded and/or assembled form, which is operative and functional. The native state of a biomolecule may possess all four levels of biomolecular structure, with the secondary through quaternary structure being formed from weak interactions along the covalently-bonded backbone. This is in contrast to the Denaturation (biochemistry), denatured state, in which these weak interactions are disrupted, leading to the loss of these forms of structure and retaining only the biomolecule's primary structure. Biochemistry Proteins While all protein molecules begin as simple unbranched chains of Amino acid, amino acids, once completed they assume highly specific three-dimensional shapes. That ultimate shape, known as tertiary structure, is the folded shape that possesses a minimum of Thermodynamic free energy, free energy. It is a protein's tertiary, folded structure that makes it capable of perform ...
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Sodium Dodecyl Sulfate
Sodium dodecyl sulfate (SDS) or sodium lauryl sulfate (SLS), sometimes written sodium laurilsulfate, is an organic compound with the formula and structure . It is an anionic surfactant used in many cleaning and hygiene products. This compound is the sodium salt of the 12-carbon organosulfate. Its hydrocarbon tail combined with a polar " headgroup" give the compound amphiphilic properties that make it useful as a detergent. SDS is also component of mixtures produced from inexpensive coconut and palm oils. SDS is a common component of many domestic cleaning, personal hygiene and cosmetic, pharmaceutical, and food products, as well as of industrial and commercial cleaning and product formulations. Physicochemical properties The critical micelle concentration (CMC) in water at 25 °C is 8.2 mM, and the aggregation number at this concentration is usually considered to be about 62. The micelle ionization fraction (α) is around 0.3 (or 30%). Applications Cleaning and hygie ...
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Disulfide Reduction By DTT-2
In chemistry, a disulfide (or disulphide in British English) is a compound containing a functional group or the anion. The linkage is also called an SS-bond or sometimes a disulfide bridge and usually derived from two thiol groups. In inorganic chemistry, the anion appears in a few rare minerals, but the functional group has tremendous importance in biochemistry. Disulfide bridges formed between thiol groups in two cysteine residues are an important component of the tertiary and quaternary structure of proteins. Compounds of the form are usually called ''persulfides'' instead. Organic disulfides Structure Disulfides have a C–S–S–C dihedral angle approaching 90°. The S–S bond length is 2.03 Å in diphenyl disulfide, similar to that in elemental sulfur. Disulfides are usually symmetric but they can also be unsymmetric. Symmetrical disulfides are compounds of the formula . Most disulfides encountered in organosulfur chemistry are symmetrical disulfides. Unsymm ...
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Oligonucleotides
Oligonucleotides are short DNA or RNA molecules, oligomers, that have a wide range of applications in genetic testing, research, and forensics. Commonly made in the laboratory by solid-phase chemical synthesis, these small fragments of nucleic acids can be manufactured as single-stranded molecules with any user-specified sequence, and so are vital for artificial gene synthesis, polymerase chain reaction (PCR), DNA sequencing, molecular cloning and as molecular probes. In nature, oligonucleotides are usually found as small RNA molecules that function in the regulation of gene expression (e.g. microRNA), or are degradation intermediates derived from the breakdown of larger nucleic acid molecules. Oligonucleotides are characterized by the sequence of nucleotide residues that make up the entire molecule. The length of the oligonucleotide is usually denoted by " -mer" (from Greek ''meros'', "part"). For example, an oligonucleotide of six nucleotides (nt) is a hexamer, while one of 25 ...
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Organelle
In cell biology, an organelle is a specialized subunit, usually within a cell (biology), cell, that has a specific function. The name ''organelle'' comes from the idea that these structures are parts of cells, as Organ (anatomy), organs are to the Human body, body, hence ''organelle,'' the suffix ''-elle'' being a diminutive. Organelles are either separately enclosed within their own lipid bilayers (also called membrane-bounded organelles) or are spatially distinct functional units without a surrounding lipid bilayer (non-membrane bounded organelles). Although most organelles are functional units within cells, some functional units that extend outside of cells are often termed organelles, such as cilia, the flagellum and archaellum, and the trichocyst (these could be referred to as membrane bound in the sense that they are attached to (or bound to) the membrane). Organelles are identified by microscopy, and can also be purified by cell fractionation. There are many types of organe ...
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Centrifuge
A centrifuge is a device that uses centrifugal force to subject a specimen to a specified constant force - for example, to separate various components of a fluid. This is achieved by spinning the fluid at high speed within a container, thereby separating fluids of different densities (e.g. cream from milk) or liquids from solids. It works by causing denser substances and particles to move outward in the radial direction. At the same time, objects that are less dense are displaced and moved to the centre. In a laboratory centrifuge that uses sample tubes, the radial acceleration causes denser particles to settle to the bottom of the tube, while low-density substances rise to the top. A centrifuge can be a very effective filter that separates contaminants from the main body of fluid. Industrial scale centrifuges are commonly used in manufacturing and waste processing to sediment suspended solids, or to separate immiscible liquids. An example is the cream separator found in da ...
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Sonicator
A sonicator at the Weizmann Institute of Science during sonicationSonication is the act of applying sound energy to agitate particles in a sample, for various purposes such as the extraction of multiple compounds from plants, microalgae and seaweeds. Ultrasonic frequencies (> 20 kHz) are usually used, leading to the process also being known as ultrasonication or ultra-sonication. In the laboratory, it is usually applied using an ''ultrasonic bath'' or an ''ultrasonic probe'', colloquially known as a ''sonicator''. In a paper machine, an ultrasonic foil can distribute cellulose fibres more uniformly and strengthen the paper. Effects Sonication has numerous effects, both chemical and physical. The scientific field concerned with understanding the effect of sonic waves on chemical systems is called sonochemistry. The chemical effects of ultrasound do not come from a direct interaction with molecular species. Studies have shown that no direct coupling of the acoustic f ...
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Homogenizer
A homogenizer is a laboratory or industrial device used to break down and evenly distribute particles within a liquid mixture, creating a stable and uniform emulsion, suspension, or solution. Homogenization (chemistry), Homogenization is a key process in many fields, including food and beverage production, pharmaceuticals, biotechnology, and materials science. It is used to process substances such as tissue, cells, soil, plant matter, and emulsified products like creams, lotions, or milk. Applications Homogenizers are widely used in both laboratory research and commercial manufacturing. Common applications include: * Cell disruption for DNA, RNA, and protein extraction * Food and beverage production (e.g., milk, juices, sauces) * Cosmetic product formulation (e.g., creams, gels, lotions) * Pharmaceutical suspensions and emulsions * Soil and plant sample processing for environmental testing Types of homogenizers A variety of technologies are used in homogenization, each with a ...
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Blender
A blender (sometimes called a mixer (from Latin ''mixus, the PPP of miscere eng. to Mix)'' or liquidiser in British English) is a kitchen and laboratory appliance used to mix, crush, purée or emulsify food and other substances. A stationary blender consists of a blender container with a rotating metal or plastic blade at the bottom, powered by an electric motor that is in the base. Some powerful models can also crush ice and other frozen foods. The newer immersion blender configuration has a motor on top connected by a shaft to a rotating blade at the bottom, which can be used with any container. Characteristics Different blenders have different functions and features but product testing indicates that many blenders, even the less expensive ones, are useful for meeting many consumer needs. Features which consumers consider when purchasing a blender include the following: *large visible measurement marks *ease of use *low noise during usage *power usage (typically 300–1000 ...
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Proceedings Of The National Academy Of Sciences
''Proceedings of the National Academy of Sciences of the United States of America'' (often abbreviated ''PNAS'' or ''PNAS USA'') is a peer-reviewed multidisciplinary scientific journal. It is the official journal of the National Academy of Sciences, published since 1915, and publishes original research, scientific reviews, commentaries, and letters. According to ''Journal Citation Reports'', the journal has a 2022 impact factor of 9.4. ''PNAS'' is the second most cited scientific journal, with more than 1.9 million cumulative citations from 2008 to 2018. In the past, ''PNAS'' has been described variously as "prestigious", "sedate", "renowned" and "high impact". ''PNAS'' is a delayed open-access journal, with an embargo period of six months that can be bypassed for an author fee ( hybrid open access). Since September 2017, open access articles are published under a Creative Commons license. Since January 2019, ''PNAS'' has been online-only, although print issues are available ...
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Surfactants
Surfactants are chemical compounds that decrease the surface tension or interfacial tension between two liquids, a liquid and a gas, or a liquid and a solid. The word ''surfactant'' is a blend of "surface-active agent", coined in 1950. As they consist of a water-repellent and a water-attracting part, they enable water and oil to mix; they can form foam and facilitate the detachment of dirt. Surfactants are among the most widespread and commercially important chemicals. Private households as well as many industries use them in large quantities as detergents and cleaning agents, but also for example as emulsifiers, wetting agents, foaming agents, antistatic additives, or dispersants. Surfactants occur naturally in traditional plant-based detergents, e.g. horse chestnuts or soap nuts; they can also be found in the secretions of some caterpillars. Today one of the most commonly used anionic surfactants, linear alkylbenzene sulfates (LAS), are produced from petroleum ...
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