The improved mechanical characteristics of this fast flow medium allows high flow. Imac sepharose 6 fast flow in buffers containing reducing agents when not in use. Ge healthcare life sciences 17531806 ni sepharose 6 fast flow affinity tag media, 5 ml volume. Ge healthcare ni sepharose 6 fast flow life sciences.
Heparin sepharose 6 fast flow hiprep heparin ff 1610 fig 1. Deae sepharose is a fast flow, weak anion exchanger suitable for the initial purification of ribonucleoproteins from cell crude extracts. Nhsactivated sepharose 4 fast flow vwr international. Substances are separated on the basis of their varying strength of their hydrophobic interaction with. Ge healthcare lifescience ni sepharose 6 fast flow is part of a range of imac chromatography media for capture and intermediate. Chromatography medium characteristics ni sepharose 6 fast flow consists of 90. Ni sepharose 6 fast flow is supplied preswollen in 5 ml, 25 ml, 100 ml, 500 ml, 1 l and, 5 l packs. Ni sepharose 6 fast flow ff, his mag sepharose ni, which are precharged with nickel. Ge healthcare life sciences 17531806 ni sepharose 6 fast. Ni sepharose 6 fast flow pdf ni sepharose 6 fast flow pdf ni sepharose 6 fast flow pdf download. Firefighting measures promptly isolate the scene by removing all persons from the vicinity of the incident if there is a fire.
Sepharose 6 fast flow high sub, phenyl sepharose 6 fast flow low sub, phenyl sepharose high performance, butyl sepharose 4 fast flow, butyls sepharose 6 fast flow and octyl sepharose 4 fast flow, code number 11003453. Ge healthcare butyls sepharose 6 fast flow chromatography media. Ge healthcare nprotein a sepharose 4 fast flow affinity media. Ni sepharose medium, provides fast, simple, and easy separations in a convenient format. Maintain packing flow rate for at least 3 bed volumes after a constant. What is the best way of packing sepharose cl6b matrix in a column. Cm sepharose fast flow is part of the sepharose fast flow ion exchange platform, which has been the industrial standard for ion exchange chromatography during recent decades. Purification of globular actin from rabbit muscle and pyrene fluorescent assays to investigate actin dynamics in vitro. Nhsactivated sepharose 4 fast flow is available in a range of different bulk pack sizes and convenient prepacked formats for easy scaleup and process development. Ni sepharose 6 fast flow ge healthcare, 17531806, pack. Four types of packing methods suitable for q sepharose fast flow are listed below. Cm sepharose fast flow ge healthcare, 17071910, pack of 25. Recombinant protein g is produced in a strain of e. Chelating sepharose fast flow consists of iminodiacetic acid groups coupled to sepharose 6 fast flow by stable ether linkages via a 7atom spacer.
The titration curve in figure 3 shows the ph working range of deae sepharose fast flow, i. Ni sepharose 6 fast flow is a high bindingcapacity imac chromatography resin for purifying histagged proteins. This prepacked column is ideal for preparative purification of histidinetagged recombinant proteins by immobilized metal ion affinity chromatography imac. Chelating sepharose fast flow consists of iminodiacetic acid groups coupled to sepharose 6 fast flow by stable ether linkages and sufficiently long spacer arms fig 2. Phenyl sepharose 6 fast flow is composed of crosslinked, 6 % agarose beads modified with standard aromatic phenyl groups via uncharged, chemicallystable ether linkages.
Phenyl sepharose 6 fast flow is part of the sepharose fast flow hic platform, which has been an industrial standard for hic chromatography during recent decades. Ni sepharose 6 fast flow consists of 90 m beads of highly crosslinked agarose, to which a chelating ligand has been immobilized and subsequently charged. Sepharose fast flow ion exchangers are easy to pack since their rigidity allows the use of high flow rates, see figure 2. Weve previously used ni nta and ni sepharose 6 fast flow resin for purification and have only seen slightly better results in favor of the ni sepharose. Ge healthcares ni sepharose 6 fast flow biocompare. Q ff is prepacked with q sepharose fast flow, and is a strong anion exchanger for smallscale protein purifications as well as screening of binding and elutions conditions. Immobilized metal ion affinity chromatography imac, developed by porath 1975, is based on the interaction of certain protein residues histidines, cysteines, and to some extent tryptophans with cations of transition metals. Its high flow properties make it excellent for scaleup. The binding capacity of protein g sepharose 4 fast flow depends on the source of the particular immunoglobulin, see table 3. This product is prepared by covalently coupling highly purified recprotein g to sepharose 4b. Data file 18101291 ac affinity chromatography protein g. Good choice for purification at a variety of scales. Column packing imac sepharose 6 fast flow is supplied preswollen in 20% ethanol. For ni sepharose 6 fast flow, very low leakage was seen over a wide interval of nickel capacities tested, demonstrating that.
The improved mechanical characteristics of this fast flow medium allows high flow rates to be used for rapid and. Total solutions for preparation of histidinetagged proteins. Immobilized metal ion affinity chromatography imac affiland. Note increasing levels of endotoxin associated with late eluting peaks. Blue sepharose 6 fast flow blue sepharose 6 fast flow is cibacron blue 3g covalently attached to the matrix by the triazine coupling method, giving a highly stable medium with minimal nonspeci. Prepare a slurry by decanting the 20% ethanol solution and replacing it with distilled water in a ratio of 75% settled medium to 25% distilled water. Pdf imac sepharose 6 fast flow, hiprep imac ff 1610, hitrap. Data file 18117288 ab ion exhange chromatography q. The molecular weight for the dye as a sodium salt is 726. Full user instructions for packing, optimization, operation, cleaning, and recharging are supplied with each pack.
Regulatory support files rsf are available to assist process validation and submissions to regulatory authorities. Histrap ff, 1 ml and 5 ml histrap ff is a ready to use column, prepacked with precharged ni sepharose 6 fast flow. Ni sepharose 6 fast flow histidinetagged protein purification resin, 500 ml cytiva. This make ni sepharose high performance the firstchoice medium for. The base matrix, sepharose 6 fast flow, is a highly crosslinked, 6 % agarose derivative with excellent kinetics, making them ideal for process scale applications, particularly during initial capture and intermediate stages of a separation process, when high flow rates are required. Ni sepharose 6 fast flow is available in 5, 25, 100, 500 ml, 1l, and 5l bulk packs.
Hiprep heparin ff 1610 is a prepacked, ready to use 20 ml column for affinity chromatography. Imac sepharose 6 fast flow is a bioprocess medium that consists of 90m beads of highly crosslinked agarose to which a chelating group has been covalently coupled. This make ni sepharose high performance the firstchoice medium for high performance purification of histidinetagged proteins. Ni sepharose 6 fast flow consists of a metal chelating moiety coupled to highly crosslinked 90 um agarose beads.
Ge healthcare ni sepharose 6 fast flow combines the advantages of ni2 for purification of histidine tagged proteins, with the well industrially established properties of the sepharose fast flow platform. Ni sepharose 6 fast flow is available in a range of different bulk pack sizes and convenient prepacked formats for easy scaleup and process development. Pdf organic solvents determination in cnbr sepharose cl4b. Ni sepharose and imac sepharose are the latest additions to the ge healthcare range of convenient and reliable solutions for purifying histidinetagged proteins. Ni sepharose 6 fast flow consists of 90 m beads of highly crosslinked agarose, to which a chelating ligand has been immobilized.
As a general method we recommend 20 mm sodium phosphate, ph 7. Ge healthcare ni sepharose 6 fast flow fisher scientific. Ge healthcare lifescience ni sepharose 6 fast flow is part of a range of imac chromatography media for capture and intermediate purification. Selecting ni sepharose products for manualautomated purification. In either case the sample is passed over the column at 4 c using the following fplc protocol. As member of the bioprocess media range, nhsactivated sepharose 4 fast flow meets industrial demands with security of supply and comprehensive technical and regulatory support. Data file 18117141 ab affinity chromatography ge healthcare life sciences chelating sepharose fast flow chelating sepharose fast flow is a bioprocess medium resin for immobilized metal ion affinity chromatography imac of native and histidinetagged fusion proteins fig 1. Data file 18117141 ab affinity chromatography chelating. Heparin sepharose 6 fast flow is available as process packs, lab packs, and prepacked hiprep heparin ff 1610. Sp sepharose fast flow, q sepharose fast flow, deae sepharose fast flow, cm sepharose fast flow, anx sepharose 4 fast flow high sub, sp sepharose xl and q sepharose xl.
Oct 01, 2009 understanding the mechanism of virus removal by q sepharose fast flow chromatography during the purification of chocell derived biotherapeutics. Ge healthcare sepharose fast flow cation exchange media cm. If the volume of the crude extract is larger than 5 mls use the 200 ml deae column, otherwise use the 30 ml deae column. Protein z chromatographed on macroprep high q with overlay of endotoxin per mg of protein z in pooled fractions. Sp sepharose fast flow is a sulphopropyl sp strong cation exchange chromatography resin for fast protein purification. Used for purification of monoclonal and polyclonal antibodies. Proteins and other molecules containing primary amino groups can be coupled. Ni sepharose 6 fast flow is a bioprocess medium that combines the advantages of ni for purification of histidine tagged proteins, with the well industrially established properties of the sepharose fast flow platform. The terminal carboxyl group is activated by esterification with nhydroxysuccinimide. Ge healthcare butyls sepharose 6 fast flow chromatography media the least hydrophobic medium in the sepharose fast flow hydrophobic interaction chromatography hic range and is particularly useful during the initial stages of purification process. As member of the bioprocess media range, ni sepharose 6 fast flow meets industrial demands with security of supply and comprehensive technical and regulatory support. Only at the end i allow for some flow, usually just by the gravity, through the column to pack the bead a bit,i insert the. By coupling heparin to sepharose 6 fast flow, proteins and other biomolecules. No action shall be taken involving any personal risk or without suitable training.
Purifying histidinetagged recombinant proteins by immobilized. Proteins with histidine tag can be purified and detected easily because the string of histidine residues binds to several types o. Hiload q and sp sepharose high performance column characteristics. What is the best way of packing sepharose cl6b matrix in a. Ge healthcare data file 18101291 ac affinity chromatography protein g sepharose 4 fast flow protein g sepharose 4 fast flow is recombinant protein g coupled to sepharose 4 fast flow. What is the different between nickel sepharose excel and. Combines the advantages of ni2 for purification of histidine tagged proteins, with the well industrially established properties of the sepharose fast flow platform. Ni sepharose 6 fast flow from ge healthcare selectscience.
Nhsactivated sepharose 4 fast flow is a beadformed, highly crosslinked preactivated matrix prepared by coupling sepharose 4 fast flow with 6 aminohexanoic acid via a spacer arm. Ni sepharose and imac sepharose hebrew university of. Designed for scaling up purification of histidinetagged proteins. Strauss dm1, lute s, tebaykina z, frey dd, ho c, blank gs, brorson k, chen q, yang b. The crosslinking of the matrix has been optimized to give. This resin is used to purify histidinetagged histagged. Igg sepharose 6 fast flow lgg sepharosetm 6 fast flow, from ge healthcare, is based on the rigid sepharose 6 fast flow matrix, with human igg covalently coupled to it. Pdf heparin sepharose 6 fast flow, hiprep heparin ff 1610. Ni sepharose 6 fast flow, 25 ml cytiva formerly ge. Any customer that wishes to use chelating sepharose fast flow, ni sepharose 6 fast flow, or imac sepharose 6 fast flow for nonresearch commercial applications under these patents is requested to contact hoffmanla roche ag, corporate licensing, attention dr. The coupling chemistry and the novel ligand is designed to give minimal ligand and ni2. Ni sepharose 6 fast flow histidinetagged protein purification resin. Ge healthcare ni sepharose 6 fast flow vol 25ml ge.
Phenyl sepharose 6 fast flow low sub, phenyl sepharose 6 fast flow high sub, butyl sepharose 4 fast flow and octyl sepharose 4 fast flow are media for hydrophobic interaction chromatography hic. Cm sepharose fast flow is a weak cation exchanger based on the well established sepharose fast flow ion exchange platform, extensively used for preparative protein separations in both research and industrial applications. Standard aromatic hic media based on sepharose fast flow base matrix derivatized via uncharged, chemicallystable ether linkages. Phenyl sepharose 6 fast flow is a well established, standard aromatic hydrophobic interaction chromatography hic medium for capture and intermediate purification requiring low to medium hydrophobicity. The sepharose 6 fast flow base matrix is a highly crosslinked, 6 % agarose with excellent chemical and physical stability. Cnbractivated sepharose 4 fast flow is a beadformed, highly crosslinked preactivated matrix produced by reacting sepharose 4 fast flow with cyanogen bromide cnbr. A slurry of 500 ml of deae sepharose fast flow beads amersham pharmacia biotech, piscataway, nj is degassed under vacuum for 30 min and packed in a column at 4. The medium is available in 25 ml, 100 ml, 1 l and 5 l packs, as well as prepacked in 1 ml and 5 ml hitrap columns and 20 ml hiprep 1610 columns. Phenyl sepharose 6 fast flow high sub cytiva formerly. Combined pressuresuction packing for medium sized columns with fixed bed heights. This resin is used to purify histidinetagged histagged and untagged recombinant proteins even at large scales.
Understanding the mechanism of virus removal by q sepharose. Butyl sepharosetm 4 fast flow octyl sepharose 4 fast flow. This coupling makes the medium more rigid which in turn improves the pressure flow characteristics. Ni sepharose 6 fast flow is a high bindingcapacity imac chromatography resin for. Download fulltext pdf organic solvents determination in cnbr sepharose cl4b and immunoaffinity matrix. Column packing guidelines general purifying biological macromolecules by hic is a typical high selectivity. Ni sepharose 6 fast flow is supplied preswollen in 5 ml, 25 ml. Imac sepharose 6 fast flow imac sepharose 6 fast flow, hitrap imac ff, hiprep imac ff 1610uncharged medium and prepacked columns for scaleup imac sepharose 6 fast flow is an uncharged medium for screening for optimal purification conditions and scaling up purification of histidinetagged and untagged proteins. Instructions chelating sepharose fast flow the wolfson. Ni sepharose 6 fast flow is composed of crosslinked 6 % agarose beads modified with a novel chelating ligand immobilized to the base matrix and precharged with ni2. The medium is easy to pack in a wide range of columns table 2. Search results for ni sepharose 6 fast flow at sigmaaldrich.
Igg from most species binds protein g sepharose 4 fast flow at neutral ph and physiological ionic strength. Imac sepharose 6 fast flow is a related affinity medium, supplied free of metal ions, allowing the user to optimize selectivity by charging the medium with the most appropriate metal ion see data file 28404106 for more details. Sepharosetm high performance or ni sepharose 6 fast flow for working with. Q, sp, deae, and cm sepharose fast flow are based on a robust, 6% highly crosslinked beaded agarose matrix with good flow. Chelating sepharose fast flow belongs to the bioprocess media family. Data file 1886 af affinity chromatography ni sepharose. Move containers from fire area if this can be done without risk. Ni sepharose 6 fast flow, 100 ml 17531802 section 5. Available as bulk resin and as prepacked columns for use with manual. Phenyl sepharose 6 fast flow low sub phenyl sepharose 6. The blue dye binds many proteins, such as albumin, interferon, lipoproteins and blood coagulation factors. It is composed of crosslinked 6% agarose beads, with carboxymethyl cm weak cation exchange groups. Cm sepharose fast flow deae sepharose fast flow q sepharose. A m e r s h a m b i o s c i e n c e s q sepharose fast flow.
Maintain packing flow rate for at least 3 bed volumes after a constant bed height is. Ni sepharose 6 fast flow is a member of the ge healthcare range of imac chromatography media for capture and intermediate. Ge healthcare lifescience nprotein a sepharose 4 fast flow affinity media contains native protein a coupled to sepharose 4 fast flow. Application examples include highyield capture, polishing monoclonal antibodies, removing truncated species from fulllength forms, separating active from inactive forms, and clearing of viruses. Ge healthcares ni sepharose 6 fast flow biocompare product.
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