Proteins and other molecules containing primary amino groups can be coupled. Wellproven strong anion exchanger developed for industrial downstream processes. Ge healthcare butyl sepharose 4 fast flow chromatography. 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. Click on the chromatography method you are interested in to jump to the corresponding selection guide. 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. Cterminal cysteine favours oriented thioether coupling. 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. Column packing imac sepharose 6 fast flow is supplied preswollen in 20% ethanol.
Adsorption performance of proteins to cm sepharose ff and. Blue sepharose 6 fast flow 50 ml 17094801 blue sepharose 6 fast flow 500 ml 17094802. The charged group of qsepharose is a quarternary amine which carries a nontitratable positive charge. 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 sepharose 6 fast flow base matrix is a highly crosslinked, 6% agarose with excellent chemical and physical stability. Deae, cm or anx if the selectivity of q or s is unsatisfactory.
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. Cm sepharose fast flow ge healthcare life sciences. This matrix can be used at alkaline ph values at which. 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. Cm sepharose fast flow deae sepharose fast flow q sepharose. Other factors such as sample stability, scale, speed, binding capacity, and. Mechanical characterisation of agarosebased chromatography. Data file 18117288 ab ion exhange chromatography q.
It is composed of crosslinked 6% agarose beads, with diethylaminoethyl deae. Sp, cm, q, and deae are based on a robust, 6% highly crosslinked beaded agarose matrix with excellent flow properties and high loading capacity. 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. In its preactivated cnbr form, it offers much improved performance when compared with the well established cnbractivated sepharose 4b.
What is the difference between sephadex and sepharose. Ideally, sepharose 4 fast flow based media are packed at a constant pressure of approximately 1 bar 0. 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. Ion exchange chromatography columns and media selection. Ge healthcare sepharose fast flow cation exchange media. Sepharose is a tradename for a crosslinked, beadedform of agarose, a polysaccharide polymer material extracted from seaweed. Typical pressureflow rate curves for butyl sepharose 4 fast flow in an xk 5030 column, bed height 15 cm. Ion exchange chromatography columns and resins ge healthcare. Deae sepharose fast flow weak anion exchanger phenyltoyopearl 630s hydrophobic matrix superdex200 gel filtration the additional set includes. Pdf q sepharose high performance and sp sepharose high. Two strong exchangers are q sepharose fast flow and sp sepharose fast flow. 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. Instructions 71500015 af preactivated media cnbractivated.
Q, sp, deae, and cm sepharose fast flow are based on a robust, 6% highly. If the recommended pressure or flow rate cannot be obtained, use. Data file 18117141 ab affinity chromatography chelating. 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. A m e r s h a m b i o s c i e n c e s q sepharose fast flow. 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. Pdf heparin sepharose 6 fast flow, hiprep heparin ff 16. Deaesepharose fast flow weak anion exchanger phenyltoyopearl 630s hydrophobic matrix superdex200 gel filtration the additional set includes. Sepharose fast flow ion exchangers include media that are called weak cm, deae, and anx or strong sp and q. Oh s rprotein a rprotein a sepharose fa st flow protein a sepharose 4 fast flow. The charged group of q sepharose is a quarternary amine which carries a nontitratable positive charge. Property description ion exchange type strong anion t otal ionic capacity 0.
Guide to ionexchange chromatography harvard apparatus. Iex sepharose fast flow labpacks and prepacked columns. Q sepharose high performance and sepharose high sp sepharose high performance q sepharose high performance, and sp sepharose high performance, are strong ion. Imac sepharose 6 fast flow in buffers containing reducing agents when not in use. Instructions71500964ag ionexchangeresins cmsepharose. It is composed of crosslinked 6% agarose beads, with diethylaminoethyl deae weak anion exchange groups. General description preswollen in 20% ethanol ccf100500ml.
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. Cm sepharose fast flow cytiva, formerly ge healthcare life. Chelating sepharose fast flow purifies proteins and peptides. Instructions71500964ag ionexchangeresins cmsepharosefastflow deaesepharosefastflow qsepharosefastflow spsepharosefastflow cm,deae. Q sepharose fast flow strong anion exchanger cm sepharose fast flow weak cation exchanger deaetoyopearl 650s weak anion exchanger sptoyopearl 650s strong cation exchanger. Data file 18117722 ab ion exchange chromatography iex. Guide to ionexchange chromatography 3 introduction cont. 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. Ge healthcare butyl sepharose 4 fast flow chromatography media. There is different type of sepharose, best of all will be sp sepharose high performance, which has particle size 24 m44 m while sp. This coupling makes the medium more rigid which in turn improves the pressureflow characteristics. Ideally, sepharose 4 fast flow media are packed in xk or tricorn columns in a twostep procedure. Its brand name is a portmanteau derived from separationpharmaciaagarose.
The ion exchange group is a carboxy methyl group, see below. A typical pressureflow rate curve for sepharose fast flow ion exchangers. These agarosebased chromatography resins have an average particle size of 80. A 14cm bed of ge zinc imac sepharose tm fast flow media was packed in an 80 cm diameter verdot ips. 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. Material cm sepharose fast flow, deae sepharose fast flow were all.
Sepharose cl, sepharose fast flow, and sepharose high. Phenyl sepharose 6 fast flow low sub phenyl sepharose 6. A typical pressure flow rate curve for sepharose fast flow ion exchangers. 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. Pdf heparin sepharose 6 fast flow, hiprep heparin ff 1610. Hiload q and sp sepharose high performance column characteristics.
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. Gelatin sepharose gelatin sepharose 4b is gelatin coupled to sepharose 4b by the cyanogen bromide method. Wash the medium with at least 5 bed volumes tst prior to use in order. The total igg capacity of protein g sepharose 4 fast flow, under defined conditions 1, for various species. Deaesepharosefastflow qsepharosefastflow spsepharosefastflow. 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. The instructions that follow are based upon packing sepharose. For applications where extremely low leakage during purification is critical, leakage can be even further reduced by performing a blank run see page 11. Fast flow ion exchange media in the recommended xk 1620. Ge healthcare your guide to chromatography media pure proteins today. The base matrix, sepharose 6 fast flow, is a 6% highly crosslinked beaded agarose with good flow properties and high loading capacity. Select the ion exchange medium according to the objective. 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. Ge healthcare sepharose fast flow cation exchange media cm.
Cm sepharose hase been used to study diet supplements and to characterize biochemical and enzymatic properties of viperidae venom. 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. Application cm sepharose is used in affinity chromatography, protein chromatography and ion exchange chromatography. Sepharose 4 fast flow matrix sepharose 4 fast flow is a highly crosslinked agarose matrix. Instructions71500964ag ionexchangeresins cmsepharose fastflow. Sp sepharose fast flow cation exchange chromatography resin.
Cm sepharose fast flow is a weak cation exchanger based on the well established sepharose fast flow ion exchange platform, extensively used for preparative. Deae sepharose is a fast flow, weak anion exchanger suitable for the initial purification of ribonucleoproteins from cell crude extracts. The crosslinking of the matrix has been optimized to give. Butyl sepharosetm 4 fast flow octyl sepharose 4 fast flow. Two strong exchangers are qsepharose fast flow and spsepharose fast flow.
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. Characteristics of cm sepharose fast flow cm sepharose fast flow is a weak cation exchanger. 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. A common application for the material is in chromatographic separations of. Cnbractivated sepharose 4 fast flow is a beadformed, highly crosslinked preactivated matrix produced by reacting sepharose 4 fast flow with cyanogen bromide cnbr. Recommended labscale columns for chelating sepharose fast flow packing flow rate mlmin max.
Deae sepharose fast flow ion exchange chromatography media. 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. Sepharose 4 fast flow native protein a, cnbr coupled and for competitor b. 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. The plate number is calculated as in figure 1, using the formula. Chelating sepharose fast flow is supplied preswollen in 20% ethanol. Pack the igg sepharose 6 fast flow suspension in any ge healthcare column of suitable size. Sp sepharose fast flow is a sulphopropyl sp strong cation exchange chromatography resin for fast protein purification. Use hitrap columns prepacked with sepharose high performance, sepharose xl and sepharose fast flow for media selection and ph scouting. This make ni sepharose high performance the firstchoice medium for high performance purification of histidinetagged proteins. Generally sepharose is the result of evolution of sephadex.
Ion exchange chromatography columns and media selection guide. 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. If the packing equipment does not include a pressure gauge, use a packing flow velocity of approximately 400 cmh. Ideally, sepharose 6 fast flow based media are packed at a constant pressure of approximately 1. 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. The sepharose 4 fast flow matrix has higher rigidity and can thus be run at high flow velocities see table 1. Table 1 presents the basic characteristics of chelating sepharose fast flow.
101 660 1553 626 375 1491 469 1382 998 1638 353 786 937 1474 483 129 856 290 1562 1078 950 1036 1077 725 1112 1645 461 954 77 109 1120 839 256 739 381 1176 1250 630 377 636 153