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FPLC

Index FPLC

FPLC may refer to. [1]

5 relations: Fast protein liquid chromatography, French Protestant Church of London, Systems development life cycle, Union of Congolese Patriots, University of New Hampshire School of Law.

Fast protein liquid chromatography

Fast protein liquid chromatography (FPLC), is a form of liquid chromatography that is often used to analyze or purify mixtures of proteins. As in other forms of chromatography, separation is possible because the different components of a mixture have different affinities for two materials, a moving fluid (the "mobile phase") and a porous solid (the stationary phase). In FPLC the mobile phase is an aqueous solution, or "buffer". The buffer flow rate is controlled by a positive-displacement pump and is normally kept constant, while the composition of the buffer can be varied by drawing fluids in different proportions from two or more external reservoirs. The stationary phase is a resin composed of beads, usually of cross-linked agarose, packed into a cylindrical glass or plastic column. FPLC resins are available in a wide range of bead sizes and surface ligands depending on the application. In the most common FPLC strategy, ion exchange, a resin is chosen that the protein of interest will bind to the resin by a charge interaction while in buffer A (the running buffer) but become dissociated and return to solution in buffer B (the elution buffer). A mixture containing one or more proteins of interest is dissolved in 100% buffer A and pumped into the column. The proteins of interest bind to the resin while other components are carried out in the buffer. The total flow rate of the buffer is kept constant; however, the proportion of Buffer B (the "elution" buffer) is gradually increased from 0% to 100% according to a programmed change in concentration (the "gradient"). At some point during this process each of the bound proteins dissociates and appears in the effluent. The effluent passes through two detectors which measure salt concentration (by conductivity) and protein concentration (by absorption of ultraviolet light at a wavelength of 280nm). As each protein is eluted it appears in the effluent as a "peak" in protein concentration and can be collected for further use. FPLC was developed and marketed in Sweden by Pharmacia in 1982 and was originally called fast performance liquid chromatography to contrast it with HPLC or high-performance liquid chromatography. FPLC is generally applied only to proteins; however, because of the wide choice of resins and buffers it has broad applications. In contrast to HPLC the buffer pressure used is relatively low, typically less than 5 bar, but the flow rate is relatively high, typically 1-5 ml/min. FPLC can be readily scaled from analysis of milligrams of mixtures in columns with a total volume of 5ml or less to industrial production of kilograms of purified protein in columns with volumes of many liters. When used for analysis of mixtures the effluent is usually collected in fractions of 1-5 ml which can be further analyzed, e.g. by MALDI mass spectrometry. When used for protein purification there may be only two collection containers, one for the purified product and one for waste.

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French Protestant Church of London

The French Protestant Church of London (Église protestante française de Londres) is a Reformed / Presbyterian church that caters to the French-speaking community of London since 1550.

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Systems development life cycle

Model of the systems development life cycle, highlighting the maintenance phase The systems development life cycle (SDLC), also referred to as the application development life-cycle, is a term used in systems engineering, information systems and software engineering to describe a process for planning, creating, testing, and deploying an information system.

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Union of Congolese Patriots

The Union of Congolese Patriots (Union des Patriotes congolais, or UPC) is a political and militia group in Ituri, northeastern Democratic Republic of the Congo, formed towards the end of the Second Congo War.

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University of New Hampshire School of Law

The University of New Hampshire School of Law is an American Bar Association–accredited law school located in Concord, New Hampshire, United States.

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References

[1] https://en.wikipedia.org/wiki/FPLC

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