Superflow 6

Superflow 6 beads exhibit the same high biological stability as the Uniflow media. The 6% beads are further stabilized by a chemical hardening reaction that supports flow rates up to 5 times higher than the regular crosslinked 4% beads. Superflow 6 is an excellent choice as a support for immobilization or for gel filtration.
The media are stable in organic solvents and aqueous solutions in the pH range 2-14. Oxidizing materials should be avoided since they can cause partial decomposition of the polysaccharide chains. The media are also resistant to high concentrations of chaotropic agents, such as 4M KI or 4M NaSCN. They can be sterilized by autoclaving at 121 °C and pH 7 for 30 min.

Uniflow, Superflow and Ultraflow are provided as aqueous suspensions with 20% ethanol as preservative. They can be stored for years at 2-8 °C. The media should not be frozen. For packing, prepare a 50% slurry and de-gas it if necessary. Packing can be performed by gravity or pump. Make sure that the operating flow rate does not exceed the packing flow rate. Equilibrate the
column with 2-3 bed volumes of buffer. For elution, a peristaltic pump is recommended, as a consistent flow rate will aid in reproducible separations. The media can be cleaned in situ with one column volume of 0.5 M NaOH, which cleans and sanitizes the resin. Autoclaving, another sterilization method, can be done before packing the column.
After cleaning with base, the column bed should be re-equilibrated with starting buffer. Superflow is an ideal support matrix for immobilizing biologically active substances. Superflow exhibits the very low nonspecific binding. This is most important when small amounts of biologics need to be isolated by affinity methods (e.g. diagnostic tests) or when a very high degree of purity is required (injectable therapeutics).

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Product Code: 806
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Table 1. Resin Characteristics

CrosslinkerEPI Chlorohydrin
Bead Material Agarose
Bead Percentage 4%
Bead Size60-160 µm
Mean Size90 μm
Flow Rate1>20 mL/min (>680 cm/hr) @ 25oC
pH Stability, Short Term22-14
pH Stability, Long Term23-13
Storage Temperature2-8o C
Shelf Lifeyears @ 2-8 oC
Storage Buffer 20% Ethanol
FormSlurry
Chemical Stability2Stable in all commonly used aqueous solutions and buffers.
Physical Stability2Negligible volume variation due to changes in pH or ionic strength.

1Linear flow rate = volumetric flow rate (cm3/h)/column cross-sectional area (cm2)

2Data refer to the coupled product, provided that the ligand can withstand the pH or chemical environment. Please note the following: pH stability, long term refers to the pH interval where the medium is stable over a long period of time without adverse effects on its subsequent chromatographic performance. pH stability, short term refers to the pH interval for regeneration and cleaning procedures.

Superflow 6 is manufactured by Sterogene Bioseparations with our proprietary cross-linking chemistry. The beads are 6% agarose and the modifications allow the gel to withstand multiple rounds of sanitization using 1M NaOH. The resin can also tolerate chromatography using chaotropes such as 4M Guanidine SCN, 6M Guanidine HCl, or 8M Urea. The beads are 60 – 160 microns in diameter, so the resin can be used for size exclusion chromatography. It is important to note that the chemistry employed for manufacture of this resin makes its separation qualities different than for other 6% cross-linked agarose resins, so curves generated by other laboratories may not apply to Superflow 6. For this reason, we recommend running molecular weight calibration standards when accurate size determinations are required.
Buffers should be filtered with 0.2 or 0.45 micron filters to reduce clogging, and very high viscosity solutions may lead to some trailing and asymmetry of the elution profiles. For analytical methods, sample volumes should be less than 5% of the volume of the Superflow 6 and less than 2% is preferable. For desalting, or buffer exchange, sample volumes of 20 – 25% are typically satisfactory.
Column geometry will also affect column resolution. Length to diameter ratios of 20 – 100 are common, with the relatively narrow columns yielding higher resolution providing that the sample volume is reduced proportionally to the reduction in diameter. In general, slower flow rates improve resolution, but there is an optimum before diffusion effects are seen.
A useful starting buffer to use with Superflow 6 is phosphate buffered saline (PBS) at pH 7 – 7.4. However, for many applications, changes in pH, ionic strength, or the addition of chaotropes may be necessary. If detergents are needed, it is usually not recommended to try and remove the detergent and use the same column for a different application.

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Instructions for use 806 Superflow6

Safety Data Sheets

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