Mitoplast
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Mitoplast
A mitoplast is a mitochondrion that has been stripped of its outer membrane leaving the inner membrane and matrix intact. How Mitoplasts Are Most Commonly Created To begin the process, mitochondria must first be separated from cultured cells. This is typically a two step process using homogenization to release the intercellular contents and differential centrifugation to separate the mitochondria from other organelles. Once the mitochondria are isolated, mitoplasts can then be formed. Mitoplasts are most commonly formed using an apparatus called a French Press. As the mitochondria pass through the narrow valve of the French press, they experience extremely high pressures around 2000 psi that rupture the outer mitochondrial membrane. The mitoplasts are then sedimented and kept in a specific storage buffer until use. When the mitoplasts are needed, they are simply placed in a potassium chloride (KCl) incubation buffer that causes the mitochondrial matrix to swell. As a result ...
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Mitoplast Patch Clamp
A mitoplast is a mitochondrion that has been stripped of its outer membrane leaving the inner membrane and matrix intact. How Mitoplasts Are Most Commonly Created To begin the process, mitochondria must first be separated from cultured cells. This is typically a two step process using homogenization to release the intercellular contents and differential centrifugation to separate the mitochondria from other organelles. Once the mitochondria are isolated, mitoplasts can then be formed. Mitoplasts are most commonly formed using an apparatus called a French Press. As the mitochondria pass through the narrow valve of the French press, they experience extremely high pressures around 2000 psi that rupture the outer mitochondrial membrane. The mitoplasts are then sedimented and kept in a specific storage buffer until use. When the mitoplasts are needed, they are simply placed in a potassium chloride (KCl) incubation buffer that causes the mitochondrial matrix to swell. As a result ...
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Mitoplast Formation
A mitoplast is a mitochondrion that has been stripped of its outer membrane leaving the inner membrane and matrix intact. How Mitoplasts Are Most Commonly Created To begin the process, mitochondria must first be separated from cultured cells. This is typically a two step process using homogenization to release the intercellular contents and differential centrifugation to separate the mitochondria from other organelles. Once the mitochondria are isolated, mitoplasts can then be formed. Mitoplasts are most commonly formed using an apparatus called a French Press. As the mitochondria pass through the narrow valve of the French press, they experience extremely high pressures around 2000 psi that rupture the outer mitochondrial membrane. The mitoplasts are then sedimented and kept in a specific storage buffer until use. When the mitoplasts are needed, they are simply placed in a potassium chloride (KCl) incubation buffer that causes the mitochondrial matrix to swell. As a result ...
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Mitochondrion
A mitochondrion (; ) is an organelle found in the cells of most Eukaryotes, such as animals, plants and fungi. Mitochondria have a double membrane structure and use aerobic respiration to generate adenosine triphosphate (ATP), which is used throughout the cell as a source of chemical energy. They were discovered by Albert von Kölliker in 1857 in the voluntary muscles of insects. The term ''mitochondrion'' was coined by Carl Benda in 1898. The mitochondrion is popularly nicknamed the "powerhouse of the cell", a phrase coined by Philip Siekevitz in a 1957 article of the same name. Some cells in some multicellular organisms lack mitochondria (for example, mature mammalian red blood cells). A large number of unicellular organisms, such as microsporidia, parabasalids and diplomonads, have reduced or transformed their mitochondria into other structures. One eukaryote, ''Monocercomonoides'', is known to have completely lost its mitochondria, and one multicellular organism, '' ...
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Outer Mitochondrial Membrane
A mitochondrion (; ) is an organelle found in the cells of most Eukaryotes, such as animals, plants and fungi. Mitochondria have a double membrane structure and use aerobic respiration to generate adenosine triphosphate (ATP), which is used throughout the cell as a source of chemical energy. They were discovered by Albert von Kölliker in 1857 in the voluntary muscles of insects. The term ''mitochondrion'' was coined by Carl Benda in 1898. The mitochondrion is popularly nicknamed the "powerhouse of the cell", a phrase coined by Philip Siekevitz in a 1957 article of the same name. Some cells in some multicellular organisms lack mitochondria (for example, mature mammalian red blood cells). A large number of unicellular organisms, such as microsporidia, parabasalids and diplomonads, have reduced or transformed their mitochondria into other structures. One eukaryote, ''Monocercomonoides'', is known to have completely lost its mitochondria, and one multicellular organism, '' Henn ...
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Electron Transport Chain
An electron transport chain (ETC) is a series of protein complexes and other molecules that transfer electrons from electron donors to electron acceptors via redox reactions (both reduction and oxidation occurring simultaneously) and couples this electron transfer with the transfer of protons (H+ ions) across a membrane. The electrons that transferred from NADH and FADH2 to the ETC involves 4 multi-subunit large enzymes complexes and 2 mobile electron carriers. Many of the enzymes in the electron transport chain are membrane-bound. The flow of electrons through the electron transport chain is an exergonic process. The energy from the redox reactions creates an electrochemical proton gradient that drives the synthesis of adenosine triphosphate (ATP). In aerobic respiration, the flow of electrons terminates with molecular oxygen as the final electron acceptor. In anaerobic respiration, other electron acceptors are used, such as sulfate. In an electron transport chain, the redo ...
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Electrochemical Gradient
An electrochemical gradient is a gradient of electrochemical potential, usually for an ion that can move across a membrane. The gradient consists of two parts, the chemical gradient, or difference in solute concentration across a membrane, and the electrical gradient, or difference in charge across a membrane. When there are unequal concentrations of an ion across a permeable membrane, the ion will move across the membrane from the area of higher concentration to the area of lower concentration through simple diffusion. Ions also carry an electric charge that forms an electric potential across a membrane. If there is an unequal distribution of charges across the membrane, then the difference in electric potential generates a force that drives ion diffusion until the charges are balanced on both sides of the membrane. Electrochemical gradients are essential to the operation of batteries and other electrochemical cells, photosynthesis and cellular respiration, and certain other bio ...
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Patch-clamp
The patch clamp technique is a laboratory technique in electrophysiology used to study ionic currents in individual isolated living cells, tissue sections, or patches of cell membrane. The technique is especially useful in the study of excitable cells such as neurons, cardiomyocytes, muscle fibers, and pancreatic beta cells, and can also be applied to the study of bacterial ion channels in specially prepared giant spheroplasts. Patch clamping can be performed using the voltage clamp technique. In this case, the voltage across the cell membrane is controlled by the experimenter and the resulting currents are recorded. Alternatively, the current clamp technique can be used. In this case, the current passing across the membrane is controlled by the experimenter and the resulting changes in voltage are recorded, generally in the form of action potentials. Erwin Neher and Bert Sakmann developed the patch clamp in the late 1970s and early 1980s. This discovery made it possible ...
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Electrophysiology
Electrophysiology (from Greek , ''ēlektron'', "amber" etymology of "electron"">Electron#Etymology">etymology of "electron" , ''physis'', "nature, origin"; and , '' -logia'') is the branch of physiology that studies the electrical properties of biological cells and tissues. It involves measurements of voltage changes or electric current or manipulations on a wide variety of scales from single ion channel proteins to whole organs like the heart. In neuroscience, it includes measurements of the electrical activity of neurons, and, in particular, action potential activity. Recordings of large-scale electric signals from the nervous system, such as electroencephalography, may also be referred to as electrophysiological recordings. They are useful for electrodiagnosis and monitoring. Definition and scope Classical electrophysiological techniques Principle and mechanisms Electrophysiology is the branch of physiology that pertains broadly to the flow of ions (ion current) in biologi ...
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Osmotic Pressure
Osmotic pressure is the minimum pressure which needs to be applied to a solution to prevent the inward flow of its pure solvent across a semipermeable membrane. It is also defined as the measure of the tendency of a solution to take in a pure solvent by osmosis. Potential osmotic pressure is the maximum osmotic pressure that could develop in a solution if it were separated from its pure solvent by a semipermeable membrane. Osmosis occurs when two solutions containing different concentrations of solute are separated by a selectively permeable membrane. Solvent molecules pass preferentially through the membrane from the low-concentration solution to the solution with higher solute concentration. The transfer of solvent molecules will continue until equilibrium is attained. Theory and measurement Jacobus van 't Hoff found a quantitative relationship between osmotic pressure and solute concentration, expressed in the following equation: :\Pi = icRT where \Pi is osmotic p ...
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Cholesterol
Cholesterol is any of a class of certain organic molecules called lipids. It is a sterol (or modified steroid), a type of lipid. Cholesterol is biosynthesized by all animal cells and is an essential structural component of animal cell membranes. When chemically isolated, it is a yellowish crystalline solid. Cholesterol also serves as a precursor for the biosynthesis of steroid hormones, bile acid and vitamin D. Cholesterol is the principal sterol synthesized by all animals. In vertebrates, hepatic cells typically produce the greatest amounts. It is absent among prokaryotes (bacteria and archaea), although there are some exceptions, such as '' Mycoplasma'', which require cholesterol for growth. François Poulletier de la Salle first identified cholesterol in solid form in gallstones in 1769. However, it was not until 1815 that chemist Michel Eugène Chevreul named the compound "cholesterine". Etymology The word "cholesterol" comes from the Ancient Greek ''chole-'' ...
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Sonication
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 chemical effects of ultrasound are concerned with understanding the effect of sonic waves on chemical systems, this is called sonochemistry. The chemical effects of ultrasound do not come from a direct interaction with molecular species. Studies have shown that no direct coupl ...
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Digitonin
Digitonin is a steroidal saponin (saraponin) obtained from the foxglove plant ''Digitalis purpurea''. Its aglycone is digitogenin, a spirostan steroid. It has been investigated as a detergent, as it effectively water-solubilizes lipids. As such, it has several potential membrane-related applications in biochemistry, including solubilizing membrane proteins, precipitating cholesterol, and permeabilizing cell membranes. Digitonin is sometimes confused with the cardiac drugs digoxin and digitoxin; all three can be extracted from the same source. Chemical properties * Critical micelle concentration = < 0.5 mM * Average micellar weight = 70000 *
Aggregation number An aggregation number is a description of the number of molecules present in a micelle once the critical micelle concentration (CMC) has been reached. In more detail, ...
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