The same procedure will be further extended to examine the adsorption performance of papain, bsa and cellulase to adsorbents to provide an effective and general evaluation method. Blue sepharose 6 fast flow 50 ml 17094801 blue sepharose 6 fast flow 500 ml 17094802. Cnbractivated sepharose 4 fast flow is a beadformed, highly crosslinked preactivated matrix produced by reacting sepharose 4 fast flow with cyanogen bromide cnbr. Cm sepharose fast flow is a weak cation exchanger based on the well established sepharose fast flow ion exchange platform, extensively used for preparative. A m e r s h a m b i o s c i e n c e s q sepharose fast flow. Ideally, sepharose 6 fast flow based media are packed at a constant pressure of approximately 1. The instructions that follow are based upon packing sepharose. Phenyl sepharose 6 fast flow low sub phenyl sepharose 6. This make ni sepharose high performance the firstchoice medium for high performance purification of histidinetagged proteins. There is different type of sepharose, best of all will be sp sepharose high performance, which has particle size 24 m44 m while sp. Instructions 71500015 af preactivated media cnbractivated. Ge healthcare butyl sepharose 4 fast flow chromatography. Ion exchange chromatography columns and media selection guide. A typical pressure flow rate curve for sepharose fast flow ion exchangers.
Data file 18117141 ab affinity chromatography chelating. Sp, cm, q, and deae are based on a robust, 6% highly crosslinked beaded agarose matrix with excellent flow properties and high loading capacity. Other factors such as sample stability, scale, speed, binding capacity, and. Q sepharose fast flow is a strong anion exchanger based on the well established sepharose fast flow ion exchange platform, extensively used for preparative protein separations in both research and industrial applications. With their high physical and chemical stability, very high batchtobatch reproducibility, and regulatory support file backup, q sepharose high performance and sp sepharose high performance are ideal for all stages of an industrial scale operation from. Use hitrap columns prepacked with sepharose high performance, sepharose xl and sepharose fast flow for media selection and ph scouting. Select the ion exchange medium according to the objective.
Chelating sepharose fast flow purifies proteins and peptides. Ion exchange chromatography columns and resins ge healthcare. Sepharose is a tradename for a crosslinked, beadedform of agarose, a polysaccharide polymer material extracted from seaweed. Cm sepharose hase been used to study diet supplements and to characterize biochemical and enzymatic properties of viperidae venom. Instructions71500964ag ionexchangeresins cmsepharose. Sepharose ion exchange media catalog numbers dcl6b100 deae sepharose cl6b dff100 deae sepharose fast flow q1126 q sepharose fast flow q1754 q sepharose high performance ccf100 cm sepharose fast flow s1799 sp sepharose fast flow storage temperature 28 c synonyms. Sepharose cl, sepharose fast flow, and sepharose high performance ion exchange media. Ge healthcare sepharose fast flow cation exchange media cm. 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 brand. Its brand name is a portmanteau derived from separationpharmaciaagarose. Wash the medium with at least 5 bed volumes tst prior to use in order.
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. The sepharose 6 fast flow base matrix is a highly crosslinked, 6% agarose with excellent chemical and physical stability. The ion exchange group is a carboxy methyl group, see below. Wellproven strong anion exchanger developed for industrial downstream processes. This coupling makes the medium more rigid which in turn improves the pressure flow characteristics. Pack the igg sepharose 6 fast flow suspension in any ge healthcare column of suitable size. Deaesepharosefastflow qsepharosefastflow spsepharosefastflow. Butyl sepharosetm 4 fast flow octyl sepharose 4 fast flow. Gelatin sepharose gelatin sepharose 4b is gelatin coupled to sepharose 4b by the cyanogen bromide method. This coupling makes the medium more rigid which in turn improves the pressureflow characteristics. The crosslinking of the matrix has been optimized to give. Cterminal cysteine favours oriented thioether coupling. A typical pressureflow rate curve for sepharose fast flow ion exchangers.
Sepharose cl, sepharose fast flow, and sepharose high. Cm sepharose fast flow deae sepharose fast flow q sepharose. Introduction anx sepharose 4 fast flow high sub is a weak anion exchanger, designed to support the increasing need for ion exchange media with different selectivity and higher chemical stability at high ph. Data file 18117288 ab ion exhange chromatography q. Cm sepharosetm fast flow, deae sepharose fast flow, q sepharose fast flow and sp sepharose fast flow are ion exchange chromatography media with excellent flow properties and high capacity for biomolecules. Characteristics of sepharose fast flow ion exchangers cation exchangers sp sepharose fast flow cm sepharose fast flow matrix 6% highly crosslinked agarose 6% highly crosslinked agarose average particle size 90 m 90 m. 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. Sepharose 4 fast flow matrix sepharose 4 fast flow is a highly crosslinked agarose matrix. Sepharose cl4b, sepharose cl6b, sepharose 4 fast flow, sepharose 6 fast flow, q sepharose high performance, mabselect and capto q ge healthcare, uppsala, sweden were used in this study. Two strong exchangers are q sepharose fast flow and sp sepharose fast flow. Cm sepharose fast flow cytiva, formerly ge healthcare life. Deae 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. For applications where extremely low leakage during purification is critical, leakage can be even further reduced by performing a blank run see page 11.
Deae sepharose fast flow ion exchange chromatography media. Q sepharose fast flow strong anion exchanger cm sepharose fast flow weak cation exchanger deaetoyopearl 650s weak anion exchanger sptoyopearl 650s strong cation exchanger. Guide to ionexchange chromatography harvard apparatus. Gelatin binds specifically to fibronectin, which is a high molecular weight glycoprotein found on the surfaces of many cell types and present in many extracellular fluids including plasma. Chelating sepharose fast flow is supplied preswollen in 20% ethanol. Guide to ionexchange chromatography 3 introduction cont. Sepharose fast flow q and deae anion exchanger s and cm cation exchanger sepharose high performance q anion exchanger sp cation exchanger minibeads columns q anion exchanger s cation exchanger. The total igg capacity of protein g sepharose 4 fast flow, under defined conditions 1, for various species.
Sepharose fast flow ion exchange media, anx has high flow rates for processing. Property description ion exchange type strong anion t otal ionic capacity 0. Q, sp, deae, and cm sepharose fast flow are based on a robust, 6% highly. These agarosebased chromatography resins have an average particle size of 80. Ge healthcare deae sepharose fast flow chromatography. The charged group of q sepharose is a quarternary amine which carries a nontitratable positive charge. Ge healthcare sepharose fast flow cation exchange media. Two strong exchangers are qsepharose fast flow and spsepharose fast flow.
Table 1 presents the basic characteristics of chelating sepharose fast flow. Sepharose fast flow ion exchangers include media that are called weak cm, deae, and anx or strong sp and q. Q sepharose high performance and sp sepharose high performance are listed in table 1. Q sepharose high performance and sepharose high sp sepharose high performance q sepharose high performance, and sp sepharose high performance, are strong ion. The initial packing was performed with an empty column, the second following the performance of reslurrying within the column. Sepharose ion exchange media product information sheet. Qsepharose fast flow strong anion exchanger cmsepharose fast flow weak cation exchanger deaetoyopearl 650s weak anion exchanger sptoyopearl 650s strong cation exchanger. Deaesepharose fast flow weak anion exchanger phenyltoyopearl 630s hydrophobic matrix superdex200 gel filtration the additional set includes. Sepharose 4 fast flow native protein a, cnbr coupled and for competitor b.
Ideally, sepharose 4 fast flow media are packed in xk or tricorn columns in a twostep procedure. If the packing equipment does not include a pressure gauge, use a packing flow velocity of approximately 500 cmh 10 cm bed height, 25 c, low viscosity buffer. What is the difference between sephadex and sepharose. This matrix can be used at alkaline ph values at which. 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. Typical pressureflow rate curves for butyl sepharose 4 fast flow in an xk 5030 column, bed height 15 cm. Pdf q sepharose high performance and sp sepharose high. Deae sepharose fast flow weak anion exchanger phenyltoyopearl 630s hydrophobic matrix superdex200 gel filtration the additional set includes. Sepharose ion exchange media catalog numbers dcl6b100 deaesepharose cl6b dff100 deaesepharose fast flow q1126 q sepharose fast flow q1754 q sepharose high performance ccf100 cm sepharose fast flow s1799 sp sepharose fast flow storage temperature 28 c synonyms. Oh s rprotein a rprotein a sepharose fa st flow protein a sepharose 4 fast flow. Iex sepharose fast flow labpacks and prepacked columns.
General description preswollen in 20% ethanol ccf100500ml. Deae, cm or anx if the selectivity of q or s is unsatisfactory. It is composed of crosslinked 6% agarose beads, with diethylaminoethyl deae. Cm sepharose fast flow ge healthcare life sciences. The sepharose 4 fast flow matrix has higher rigidity and can thus be run at high flow velocities see table 1.
Mechanical characterisation of agarosebased chromatography. The instructions that follow are based upon packing sepharose fast flow ion exchange media in the recommended xk 1620 column. Q sepharose high performance and sp sepharose high. The charged group of qsepharose is a quarternary amine which carries a nontitratable positive charge. There is different type of sepharose, best of all will be sp sepharose high performance, which has particle size 24. The plate number is calculated as in figure 1, using the formula. Click on the chromatography method you are interested in to jump to the corresponding selection guide. Imac sepharose 6 fast flow in buffers containing reducing agents when not in use. Deae sepharose is a fast flow, weak anion exchanger suitable for the initial purification of ribonucleoproteins from cell crude extracts. The base matrix, sepharose 6 fast flow, is a 6% highly crosslinked beaded agarose with good flow properties and high loading capacity. Application cm sepharose is used in affinity chromatography, protein chromatography and ion exchange chromatography. A 14cm bed of ge zinc imac sepharose tm fast flow media was packed in an 80 cm diameter verdot ips. If the packing equipment does not include a pressure gauge, use a packing flow velocity of approximately 400 cmh. A common application for the material is in chromatographic separations of.
The heparin ligand used in heparin sepharose 6 fast flow is isolated from porcine intestinal mucosa and has a molecular weight distribution of m r 5000 to 30 000. Ge healthcare your guide to chromatography media pure proteins today. Fast flow ion exchange media in the recommended xk 1620. Ideally, sepharose 4 fast flow based media are packed at a constant pressure of approximately 1 bar 0. Sp sepharose fast flow cation exchange chromatography resin.
Ge healthcare butyl sepharose 4 fast flow chromatography media. It is composed of crosslinked 6% agarose beads, with diethylaminoethyl deae weak anion exchange groups. Adsorption performance of proteins to cm sepharose ff and. Sepharose fast flow ion exchange media comprise sp sepharose fast flow, cm sepharose fast flow, q sepharose fast flow, deae sepharose fast flow, and anx sepharose 4 fast flow high sub. Recommended labscale columns for chelating sepharose fast flow packing flow rate mlmin max. Instructions71500964ag ionexchangeresins cmsepharosefastflow deaesepharosefastflow qsepharosefastflow spsepharosefastflow cm,deae. Typical pressureflow rate curves for phenyl sepharose 6 fast flow low sub and phenyl sepharose 6 fast flow high sub in an xk 5030 column, bed height 15 cm. Instructions71500964ag ionexchangeresins cmsepharose fastflow. Characteristics of cm sepharose fast flow cm sepharose fast flow is a weak cation exchanger. Generally sepharose is the result of evolution of sephadex.
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. Data file 18117722 ab ion exchange chromatography iex. Cm sepharose fast flow, deae sepharose fast flow, q sepharose fast flow and sp sepharose fast flow are ion exchange chromatography media with excellent flow properties and high capacity for biomolecules. Pdf heparin sepharose 6 fast flow, hiprep heparin ff 16. Hiload q and sp sepharose high performance column characteristics. Column packing imac sepharose 6 fast flow is supplied preswollen in 20% ethanol. Pdf heparin sepharose 6 fast flow, hiprep heparin ff 1610. In its preactivated cnbr form, it offers much improved performance when compared with the well established cnbractivated sepharose 4b. Based on sepharose fast flow base matrix derivatized via uncharged, chemicallystable ether linkages hydrophobic interaction chromatography hic media composed of crosslinked 4% agarose aliphatic butyl ligand immobilized to base matrix with ether linkage. 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. Sp sepharose fast flow is a sulphopropyl sp strong cation exchange chromatography resin for fast protein purification. Ion exchange chromatography columns and media selection.
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