|
Protein Purification
Protein purification is a series of processes intended to isolate one or a few proteins from a complex mixture, usually Cell biology, cells, Tissue (biology), 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 will not 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 manufacturin ... [...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 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 ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Lysozyme
Lysozyme (, 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 alpha-lactalbumin, α-lactalbumin in sequence and st ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Percoll
Percoll is a reagent consisting of colloidal silica particles used in cell biology and other laboratory settings. It was first formulated by Pertoft and colleagues, and commercialized by Pharmacia Fine Chemicals. Percoll is used for the isolation of cells, organelles, or viruses by density centrifugation. Percoll was developed from previously reported uses of colloidal silica nanoparticles coated with polysaccharides or polymers for rate zonal, isopycnic, or equilibrium centrifugal separations. Percoll itself specifically consists of polydisperse silica nanoparticles 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 Cytiva. Characteristics Percoll does not notably diffuse over time, resulting in the f ... [...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 Svedberg, 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 (biology), tissue sample is first Lysis, 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-pell ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
G-force
The g-force or gravitational force equivalent is a Specific force, mass-specific force (force per unit mass), expressed in Unit of measurement, units of standard gravity (symbol ''g'' or ''g''0, not to be confused with "g", the symbol for grams). It is used for sustained accelerations that cause a perception of weight. For example, an object at rest on Earth's surface is subject to 1 ''g'', equaling the conventional value of gravitational acceleration on Earth, about . More transient acceleration, accompanied with significant jerk (physics), jerk, is called Shock (mechanics), ''shock''. 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 force for every unit of each object's mass. The types of forces involved are transmitted through objects by interior Stress (mechanics), mechanical stresses. Gravitational acceleration is one cause of an object's accelera ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Acceleration
In mechanics, acceleration is the Rate (mathematics), rate of change of the velocity of an object with respect to time. Acceleration is one of several components of kinematics, the study of motion. Accelerations are Euclidean vector, vector quantities (in that they have Magnitude (mathematics), magnitude and Direction (geometry), 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 Direct proportionality, directly proportional to this net resulting force; * that object's mass, depending on the materials out of which it is made — magnitude is Inverse proportionality, inversely proportional to the object's mass. The International System of Units, SI unit for acceleration is metre per second squared (, \ma ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
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 ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Inertia
Inertia is the natural tendency of objects in motion to stay in motion and objects at rest to stay at rest, unless a force causes the velocity to change. It is one of the fundamental principles in classical physics, and described by Isaac Newton in his Newton%27s_laws_of_motion#First, first law of motion (also known as The Principle of Inertia). It is one of the primary manifestations of mass, one of the core quantitative properties of physical systems. Newton writes: In his 1687 work ''Philosophiæ Naturalis Principia Mathematica'', Newton defined inertia as a property: History and development Early understanding of inertial motion Joseph NeedhamProfessor John H. Lienhard points out the Mozi (book), Mozi – based on a Chinese text from the Warring States period (475–221 BCE) – as having given the first description of inertia. Before the European Renaissance, the prevailing theory of motion in western philosophy was that of Aristotle (384–322 BCE). On the surface ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Centrifugal Force
Centrifugal force is a fictitious force in Newtonian mechanics (also called an "inertial" or "pseudo" force) that appears to act on all objects when viewed in a rotating frame of reference. It appears to be directed radially away from the axis of rotation of the frame. The magnitude of the centrifugal force ''F'' on an object of mass ''m'' at the perpendicular distance ''ρ'' from the axis of a rotating frame of reference with angular velocity is F = m\omega^2 \rho. This fictitious force is often applied to 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 non–inertial reference frame such as a rotating coordinate system. The term has sometimes also been used for the '' reactive centrifugal force'', a real frame-independent Newtonian force that exists as a reaction to a centripetal force in some scenarios. History F ... [...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 ... [...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 animals are known as deoxy ... [...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 proteolysis, 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 inhibito ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
