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Protein Purification
Protein purification is a series of processes intended to isolate one or a few proteins from a complex mixture, usually cells, tissues or whole organisms. Protein purification is vital for the specification of the function, structure and interactions of the protein of interest. The purification process may separate the protein and non-protein parts of the mixture, and finally separate the desired protein from all other proteins. Ideally, to study a protein of interest, it must be separated from other components of the cell so that contaminants won't interfere in the examination of the protein of interest's structure and function. Separation of one protein from all others is typically the most laborious aspect of protein purification. Separation steps usually exploit differences in protein size, physico-chemical properties, binding affinity and biological activity. The pure result may be termed protein isolate. Purpose The protein manufacturing cost remains high and there is a g ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
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 ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Lysozyme
Lysozyme (EC 3.2.1.17, muramidase, ''N''-acetylmuramide glycanhydrolase; systematic name peptidoglycan ''N''-acetylmuramoylhydrolase) is an antimicrobial enzyme produced by animals that forms part of the innate immune system. It is a glycoside hydrolase that catalyzes the following process: : Hydrolysis of (1→4)-β-linkages between ''N''-acetylmuramic acid and ''N''-acetyl-D-glucosamine residues in a peptidoglycan and between ''N''-acetyl-D-glucosamine residues in chitodextrins Peptidoglycan is the major component of gram-positive bacterial cell wall. This hydrolysis in turn compromises the integrity of bacterial cell walls causing lysis of the bacteria. Lysozyme is abundant in secretions including tears, saliva, human milk, and mucus. It is also present in cytoplasmic granules of the macrophages and the polymorphonuclear neutrophils (PMNs). Large amounts of lysozyme can be found in egg white. C-type lysozymes are closely related to α-lactalbumin in sequence and structure ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Percoll
Percoll is a tool for efficient density separation in Cell biology that was first formulated by Pertoft and colleagues. It is used for the isolation of cells, organelles, and/or viruses by density centrifugation. Percoll consists of colloidal silica particles of 15–30 nm diameter (23% w/w in water) which have been coated with polyvinylpyrrolidone (PVP). Percoll is well suited for density gradient experiments because it possesses a low viscosity compared to alternatives, a low osmolarity, and no toxicity towards cells and their constituents. Percoll is a registered trademark of GE Healthcare. Past use in artificial reproduction Percoll was previously used in assisted reproductive technology (ART) to select sperm from semen by density gradient centrifugation, for use in techniques such as ''in vitro'' fertilization or intrauterine insemination. However, in 1996, Pharmacia sent out a letter to laboratories stating that Percoll should be used for research purposes only, no ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Sucrose Gradient Centrifugation
In biochemistry and cell biology, differential centrifugation (also known as differential velocity centrifugation) is a common procedure used to separate organelles and other sub-cellular particles based on their sedimentation rate. Although often applied in biological analysis, differential centrifugation is a general technique also suitable for crude purification of non-living suspended particles (e.g. nanoparticles, colloidal particles, viruses). In a typical case where differential centrifugation is used to analyze cell-biological phenomena (e.g. organelle distribution), a tissue sample is first lysed to break the cell membranes and release the organelles and cytosol. The lysate is then subjected to repeated centrifugations, where particles that sediment sufficiently quickly at a given centrifugal force for a given time form a compact "pellet" at the bottom of the centrifugation tube. After each centrifugation, the ''supernatant'' (non-pelleted solution) is removed from t ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
G-force
The gravitational force equivalent, or, more commonly, g-force, is a measurement of the type of force per unit mass – typically acceleration – that causes a perception of weight, with a g-force of 1 g (not gram in mass measurement) equal to the conventional value of gravitational acceleration on Earth, ''g'', of about . Since g-forces indirectly produce weight, any g-force can be described as a "weight per unit mass" (see the synonym specific weight). When the g-force is produced by the surface of one object being pushed by the surface of another object, the reaction force to this push produces an equal and opposite weight for every unit of each object's mass. The types of forces involved are transmitted through objects by interior mechanical stresses. Gravitational acceleration (except certain electromagnetic force influences) is the cause of an object's acceleration in relation to free fall. The g-force experienced by an object is due to the vector sum of all ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Acceleration
In mechanics, acceleration is the rate of change of the velocity of an object with respect to time. Accelerations are vector quantities (in that they have magnitude and direction). The orientation of an object's acceleration is given by the orientation of the ''net'' force acting on that object. The magnitude of an object's acceleration, as described by Newton's Second Law, is the combined effect of two causes: * the net balance of all external forces acting onto that object — magnitude is directly proportional to this net resulting force; * that object's mass, depending on the materials out of which it is made — magnitude is inversely proportional to the object's mass. The SI unit for acceleration is metre per second squared (, \mathrm). For example, when a vehicle starts from a standstill (zero velocity, in an inertial frame of reference) and travels in a straight line at increasing speeds, it is accelerating in the direction of travel. If the vehicle turns, an acc ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Centrifuge
A centrifuge is a device that uses centrifugal force 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 dairies. Very high speed centrifuges and ultracentrifuges able to provi ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Inertia
Inertia is the idea that an object will continue its current motion until some force causes its speed or direction to change. The term is properly understood as shorthand for "the principle of inertia" as described by Newton in his first law of motion. After some other definitions, Newton states in his first law of motion: The word "perseveres" is a direct translation from Newton's Latin. Other, less forceful terms such as "to continue" or "to remain" are commonly found in modern textbooks. The modern use follows from some changes in Newton's original mechanics (as stated in the ''Principia'') made by Euler, d'Alembert, and other Cartesians. The term inertia comes from the Latin word ''iners'', meaning idle, sluggish. The term inertia may also refer to the resistance of any physical object to a change in its velocity. This includes changes to the object's speed or direction of motion. An aspect of this property is the tendency of objects to keep moving in a straight li ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Centrifugal Force
In Newtonian mechanics, the centrifugal force is an inertial force (also called a "fictitious" or "pseudo" force) that appears to act on all objects when viewed in a rotating frame of reference. It is directed away from an axis which is parallel to the axis of rotation and passing through the coordinate system's origin. If the axis of rotation passes through the coordinate system's origin, the centrifugal force is directed radially outwards from that axis. The magnitude of centrifugal force ''F'' on an object of mass ''m'' at the distance ''r'' from the origin of a frame of reference rotating with angular velocity is: F = m\omega^2 r The concept of centrifugal force can be applied in rotating devices, such as centrifuges, centrifugal pumps, centrifugal governors, and centrifugal clutches, and in centrifugal railways, planetary orbits and banked curves, when they are analyzed in a rotating coordinate system. Confusingly, the term has sometimes also been used for the reactiv ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Centrifugation
Centrifugation is a mechanical process which involves the use of the centrifugal force to separate particles from a solution according to their size, shape, density, medium viscosity and rotor speed. The denser components of the mixture migrate away from the axis of the centrifuge, while the less dense components of the mixture migrate towards the axis. Chemists and biologists may increase the effective gravitational force of the test tube so that the precipitate (pellet) will travel quickly and fully to the bottom of the tube. The remaining liquid that lies above the precipitate is called a supernatant or supernate. There is a correlation between the size and density of a particle and the rate that the particle separates from a heterogeneous mixture, when the only force applied is that of gravity. The larger the size and the larger the density of the particles, the faster they separate from the mixture. By applying a larger effective gravitational force to the mixture, like a ce ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Deoxyribonuclease
Deoxyribonuclease (DNase, for short) refers to a group of glycoprotein endonucleases which are enzymes that catalyze the hydrolytic cleavage of phosphodiester linkages in the DNA backbone, thus degrading DNA. The role of the DNase enzyme in cells includes breaking down extracellular DNA (ecDNA) excreted by apoptosis, necrosis, and neutrophil extracellular traps (NET) of cells to help reduce inflammatory responses that otherwise are elicited. A wide variety of deoxyribonucleases are known and fall into one of two families (DNase I or DNase II), which differ in their substrate specificities, chemical mechanisms, and biological functions. Laboratory applications of DNase include purifying proteins when extracted from prokaryotic organisms. Additionally, DNase has been applied as a treatment for diseases that are caused by ecDNA in the blood plasma. Assays of DNase are emerging in the research field as well. Types The two main types of DNase found in metazoans are known as deoxyr ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Protease Inhibitor (biology)
In biology and biochemistry, protease inhibitors, or antiproteases, are molecules that inhibit the function of proteases (enzymes that aid the breakdown of proteins). Many naturally occurring protease inhibitors are proteins. In medicine, ''protease inhibitor'' is often used interchangeably with alpha 1-antitrypsin (A1AT, which is abbreviated PI for this reason). A1AT is indeed the protease inhibitor most often involved in disease, namely in alpha-1 antitrypsin deficiency. Classification Protease inhibitors may be classified either by the type of protease they inhibit, or by their mechanism of action. In 2004 Rawlings and colleagues introduced a classification of protease inhibitors based on similarities detectable at the level of amino acid sequence. This classification initially identified 48 families of inhibitors that could be grouped into 26 related superfamily (or clans) by their structure. According to the MEROPS database there are now 81 families of inhibitors. These fa ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
