Selection kit together with sp sepharose xl and q sepharose xl. J prepacked hitrap columns with sephadex and a wide variety. It is composed of crosslinked 6% agarose beads, with sulphopropyl sp strong cation exchange groups. The top and bottom frits are manufactured from porous polyethylene. Use for highresolution, smallscale protein purification. Q sepharose xl, q sepharose xl virus licensed, sp sepharose. Hitrap sp hp, 1 ml and 5 ml hitrap q hp, 1 ml and 5 ml. Carefully fill the rest of the column with distilled water to form an upward meniscus at the top and insert the flow adaptor.
Insights in understanding aggregate formation and dissociation in. Hitrap sp sepharose ff cytiva formerly ge healthcare. The method is based on the use of an ionic column and salt gradient elution buffer buffer solution plus 1 m nacl. Hitrap sp hp and hitrap q hp are prepacked, ready to use cation and anion exchange columns for method scouting, group separations, sample concentration and sample cleanup of charged biomolecules. Q and s sepharose high performance media are available prepacked in hitrap and hiload columns or in media packs. Recombinant protein g is produced in a strain of e. Guide to ionexchange chromatography harvard apparatus. Hitrap columns are available for either cation exchange or anion exchange chromatography. Pdf q sepharose high performance and sp sepharose high. The packing material is based on patented sepharose high performance hp beads.
Ion exchange chromatography question nov232006 can i use tris buffer ph 8. Aktadesign system includes preset method templates based on these prepacked columns, which further improves convenience and results. Hitrap sp ff is prepacked with sp sepharose fast flow, and is a strong cation exchanger for smallscale protein purifications as well as screening of binding and elutions conditions. Ge healthcare life sciences hitrap iex hp prepacked columns are prepacked, readytouse sepharose high performance strong cation exchange columns. The column was washed with tes buffer containing 20 % vv. Mount the adaptor directly on the column as close to the upper edge as possible. 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.
Hitrap q hp and hitrap sp hp are 1 ml and 5 ml columns made of polypropylene, which is biocompatible and noninteractive with biomolecules. Sp sepharose fast flow is supplied in suspension in 20% ethanol and 0. Fractionation of soluble proteins in escherichia coli. It is delivered with a stopper on the inlet and a snap. Protocols and tips in protein purification or how to purify protein in one day. The use of phenylsepharose for the affinity purification. This matrix can be used at alkaline ph values at which. Sp sepharose fast flow cation exchange chromatography. Selection of ion exchanger and conditions ion exchange chromatography is based on. Guide to ionexchange chromatography 3 introduction cont. Methods can be easily scaled up from columns such as hitrap q ff 1 ml, prepacked with q sepharose fast flow through to large scale columns such as. Connect an operable pump to the column for reproducible separations. Table 3 lists recommended columns available from amersham biosciences.
Can one use sp sepharose rather than mono s for protein. Sp sepharose has been used to study inhibitory proteins and peptides from the rhizomes of zingiberaceae plants as well as to study the subunit heterogeneity and molecular evolution of soybean basic 7s globulin. Hitrap sp hp and hitrap q hp provide fast, reproducible, and easy. Prepacked q and sp sepharose high performance columns by providing added speed, convenience and reproducibility, prepacked columns extend the usefulness of q and sp sepharose high performance. Pack the column at 1 mlmin 30 cmh until the bed is stabilized 20 min. Media characteristics q sepharose fast flow is a strong anion exchanger.
Whey protein and buffer solution ph 4 ph 10 were prepared before run the experiment. Pdf q sepharose high performance sp sepharose high. Anx sepharose 4 fast flow high sub is based on 4% highly crosslinked beaded agarose. Hitrap columns, sp sepharose ff, ge healthcare vwr. Hitrap sp hp and hitrap q hp columns characteristics column volumes 1 ml or 5 ml column. These are prepacked columns and ready to use with the simple preparation of a few buffers. Results show good correlation with data obtained by column chromatography. Description media properties sp sepharose high performance and q sepharose high performance are strong cation and strong anion exchangers. When required pepsinogen and the proproteinase from bacillus sp. Q, sp, deae, and cm sepharose fast flow are based on a robust, 6% highly crosslinked beaded agarose matrix with good flow properties and high loading capacities. Q sepharose high performance and sp sepharose high. Structurepoint, formerly the pca engineering software group, offers concrete design software programs updated to aci 31814 for concrete buildings, concrete structures and concrete tanks. Chromatography columns separate molecules within mixtures during gas or liquid methods. Filled with stationary phase compositions or resins, the prepacked options reduce the preparing time required.
Sp sepharose fast flow, aqueous ethanol suspension, 45. Sp sepharose fast flow can also be packed in the other column platforms such as bpg and chromaflow. This product is prepared by covalently coupling highly purified recprotein g to sepharose 4b. It is composed of crosslinked 6% agarose beads, with quaternary ammonium q strong anion exchange groups. 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. Fill the column with buffer to the rim and allow 5 bv to flow through the column. Protocols and tips in protein purification university of sheffield. Ge healthcare hitrap sp sepharose fast flow iex columns. Data file 18117288 ab ion exhange chromatography q. Inner dia bed bed column meter mm volume height cm labscale.
Continue packing the column at 480 20 kpa for 6 minutes. Q sepharose fast flow and sp sepharose fast flow are also available in the prefilled predictor 96well filter plates for fast and easy process development. For use with application for method scouting, group separations, sample concentration and sample cleanup of charged biomolecules. In an effort to simplify a complex mixture of soluble proteins from escherichia coli, methods to fractionate the samples prior to twodimensional 2d gel electrophoresis were developed.
Convenient and reproducible for fast, easy, highperformance separations. Hitrap sp ff columns convenient for easy use with a syringe, peristaltic pump, or chromatography system such as akta system. Q or sp sepharose xl q or sp capto q, s, mmc sepharose fast flow q, sp, deae, cm, anx. Pack the column at the flow rate which gives 480 20 kpa for 45 minutes. Sp sepharose fast flow ion exchange chromatography media. Sepharose sepharose is a tradename for a crosslinked, beadedform of agarose, a polysaccharide polymer material extracted from seaweed. Packing phenyl or butyl sepharose high performance in xk 1 20 column to obtain a 10 cm bed height, fill the column tube with a 75% medium slurry. Large pack sizes of the media in 2% benzyl alcohol 2% benzyl alcohol and 0. The ion exchange group is a quartenary amine group, see fig.
Sp sepharose high performance from ge healthcare life. To ensure best peformance and troublefree operation, please read these instructions before using q sepharose fast flow. Sp sepharose is used in protein chromatography, ion exchange chromatography and cation exchange media. Gloves and protective eyewear should be worn when handling samples and reagents buffers, however it is preferable that the user remove gloves prior to entering commands via the computer keyboard or mouse.
Once the column is prepared with mclabs sp sepharose big beads, apply or inject the specific sample dissolved in the starting buffer to the column. Sp 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. Sp sepharose ff average particle size 90 um is usually used in early capture step, whereas sp sepharose hp average particle size 34 um could be used in later polishing step. Characteristics of hitrap sp hp and hitrap q hp, 1 and 5 ml columns are listed in table 2. Q and sp sepharose xl can be used with most equipment available for chromatography from laboratory to production scale. These methods involve the use of deae sepharose, sp sepharose, and phenyl sepharose chromatographic columns and the fractionation of the protein mixtures based on differential anionic, cationic, and. The charged group of q sepharose is a quarternary amine which carries a nontitratable positive charge. Hepes or even acetate is better buffer components for sp or c columns. Bioprocess chromatography resins are developed and supported for production scale chromatography. A specific software version might be needed for the chosen system. The use of phenyl sepharose for the affinity purification of proteinases. Predictor sp sepharose fast flow cytiva formerly ge healthcare. Phenyl sepharose high performance butyl sepharose high. Assist software supports the predictor workflow from setup of experimental design to.
Selection guide ion exchange media will bind cation exchanger will bind anion exchanger select cation exchange at system ph. Q, sp, deae, and cm sepharose fast flow are based on a robust, 6% highly crosslinked beaded agarose matrix with good flow. 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. Hitrap sp high performance ge healthcare, 17115201. The adaptor should be adjusted down to the bed surface. Concrete design software frequently asked questions. Ion exchange chromatography question protein and proteomics. Hitrap sp hp are prepacked, readytouse sp sepharose high performance strong cation exchange columns for highresolution, smallscale protein purification. The program offers investigation of irregularly shaped, reinforced concrete column sections that may contain openings or boundary elements. The columns have an advanced design and are constructed from precision bore borosilicate glass. The instructions that follow are based upon packing sepharose fast flow ion exchange media in the recommended xk 1620 column. Sp sepharose fast flow uses the sepharose fast flow ion exchange resin, which is well established in industrial downstream processing. Bioprocess sp sepharose hp and q sepharose hp are part of the bioprocess range of resins.
Ge healthcare also sells prepacked hiprep 20ml columns with deae, sp, q sepharose. Deae sepharose is a fast flow, weak anion exchanger suitable for the initial purification of ribonucleoproteins from cell crude extracts. This strong cation exchange resin is available in the following prepacked formats. Ion exchange chromatography columns and media selection. For the basic purification protocol you need a set of 3 columns packed with deae sepharose fast flow weak anion exchanger phenyltoyopearl 630s hydrophobic matrix. Sp sepharose are cation exchange columns and q sepharose is an anion exchanger. A m e r s h a m b i o s c i e n c e s q sepharose fast flow. Reinforced concrete structural software includes programs for column design pcacolumn, beam design pcabeam, slab design pcaslab, wall design pcawall, mat design pcamats, foundation design.