Prepacked 25 mm × 7 mm i.d. HiTrap columns containing SP Sepharose FF, SP Sepharose XL and Capto S, all strong cation exchangers, were purchased from GE Healthcare Life Sciences. A prepacked 50 mm × 5 mm i.d. S HyperCel column was obtained from Pall Life Sciences. The physical properties of the resins are summarized in Table 2 . SP FF comprises an agarose base matrix functionalized with sulfonate groups attached to a short spacer arm. SP XL uses the same agarose base matrix as SP FF, but also contains covalently attached 40 kDa dextran extenders. Both the dextran and the agarose base matrix carry sulfonate functionalization, substantially expanding the volume available for adsorption within this material as compared to its non-polymer-modified counterpart but concomitantly reducing the pore space available for transport. Capto S contains similar 40 kDa extenders to those present in SP XL but the cross-linking density of the agarose base matrix is higher than for the Sepharose materials [7 ], resulting in a slightly smaller pore lumen [16 (link)]. S HyperCel is comprised of a fully cellulosic base matrix without any defined polymer modification [8 (link)], but it exhibits properties comparable to those seen in polymer-modified stationary phases [17 ], including increased capacity and enhanced uptake rates.
Capto s
Manufactured by GE Healthcare
Sourced in United States
Capto S is a chromatography media designed for the separation and purification of proteins, peptides, and other biomolecules. It features a rigid, high-capacity agarose-based matrix that provides excellent flow properties and mechanical stability. The product is suitable for use in a variety of chromatographic techniques, including ion exchange, hydrophobic interaction, and affinity chromatography.
Automatically generated - may contain errors
Lab products found in correlation
Capto q, by GE Healthcare (2 mentions)
Sp sepharose ff, by GE Healthcare (1 mentions)
S hypercel column, by Pall Corporation (1 mentions)
Mabselect sure, by GE Healthcare (1 mentions)
Akta explorer 10, by GE Healthcare (1 mentions)
Diaion hp 20, by Merck Group (1 mentions)
Akta prime plus, by GE Healthcare (1 mentions)
1260 infinity, by Agilent Technologies (1 mentions)
Sp sepharose fast flow, by GE Healthcare (1 mentions)
Sepharose cl 4b, by GE Healthcare (1 mentions)
10 ml syringe, by BD (1 mentions)
Fractogel emd so3 m, by Merck Group (1 mentions)
Ny1102500, by Merck Group (1 mentions)
Complete his tag purification resin, by Roche (1 mentions)
Mabselect sure lx, by GE Healthcare (1 mentions)
Superdex 75, by GE Healthcare (1 mentions)
6 protocols using capto s
1
Evaluation of Strong Cation Exchange Chromatography Resins
2
High-Yield Antibody Purification Protocol
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Stable cell lines with high antibody expression levels were obtained through methotrexate pressurization. The cell line with the highest antibody expression level was cultured in a 5-L bioreactor (Z310110010, Applikon, Delft, Netherlands). The cell culture supernatant was concentrated and purified using a purification system (AKTA Explorer 10, General Electric Company, Boston, Massachusetts, USA) and a three-step chromatographic method [26 ] as follows: MabSelect SuRe™ (17,543,802, General Electric Company) affinity chromatography to bind antibodies in the cell culture supernatant; Capto™ S (17,544,101, General Electric Company) cation exchange chromatography to exchange target antibodies by conferring a positive charge according to the isoelectric point of antibody; and Capto™ Q (17,531,602, General Electric Company) anion exchange chromatography to remove endotoxins and nucleic acids from the antibodies.
3
Protein Separation by Cation Exchange
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Example 3
The proteins in the dissolved solids fraction were separated and recovered by chromatography using the resin Capto S, a strong cation exchange medium supplied in a prepacked column (GE Healthcare). Three protein fractions were collected sequentially after elution with 0.1 M sodium acetate at pH 4.5 containing increasing amounts of NaCl at 0.2, 0.4 and 0.6 M. This allowed differential separation of pot ale proteins as demonstrated by SDS-PAGE analysis (
4
Purification of LMW Antimicrobial Peptide
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Strain IE-3 was grown anaerobically in serum vials at 30°C for 48 h for the maximum production of a LMW peptide. Antimicrobial compound was extracted from CFB using 2% activated Diaion HP20 (Sigma, USA) hydrophobic resin. The crude extract obtained was further purified through cation exchange (Capto S, GE Healthcare, USA) chromatography column linked to an AKTA prime plus (GE healthcare, USA), in 20 mM sodium acetate buffer (pH 4.6) and eluted with NaCl gradient (50 to 1000 mM) in binding buffer. The peptide was desalted using dialysis tube (molecular cutoff 0.5 kDa, Spectrum, USA). Approximate molecular mass of peptide was determined by gel filtration column (Sodex KW-802.5) using standard molecular weight markers as described earlier [31 (link)]. Purity was confirmed by reversed phase HPLC (10 mm × 250 mm × 150 Å) C-18 column (venusil, Agela Technologies) under isocratic flow (1.5 ml/min) of acetonitrile (20%) along with 0.1% TFA. Elution was monitored at 200–340 nm wavelength range on PDA detector and peaks were collected by fraction collector (1260 Infinity, Agilent technology, USA).
5
Extracellular Vesicle Isolation Using Size Exclusion and Ion Exchange Chromatography
6
Purification of H11, H11-HLE, Ipi-WT, and Ipi-LALAPG
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The culture supernatants which contained H11 and H11-HLE were loaded onto cOmplete™ His-Tag Purification Resin (Roche), followed by Capto S (GE Healthcare), Capto Q (GE Healthcare), and Superdex 75 (GE Healthcare). The final concentrated proteins were stored in PBS. The culture supernatants which contained Ipi-WT and Ipi-LALAPG were loaded onto MabSelect SuRe LX (GE Healthcare). The eluted proteins were neutralized with 1 M trisodium citrate. The final concentrated proteins were stored in 25 mM sodium citrate, 125 mM NaCl pH 5.5. All purified proteins were > 95% pure, confirmed by SDS-PAGE. All endotoxin levels were below 0.02 E.U./mg.
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