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