Solid Phase Extraction Tools
PolyLC offers all of its materials in cartridges as Solid Phase Extraction Tools as well as additional materials:
Solid Phase Extraction Tools NuTips® and TopTips®
NuTips®: Available for samples 1-10 µl and 10-200 µl. The stationary phase is embedded in the wall with an open channel in the center. Sample is pipetted up and down to promote solute capture. These are convenient for desalting or quick sampling and spotting on a MALDI target. However, if the amount of peptide being sampled greatly exceeds the capacity of the NuTip (or any other SPE cartridge, for that matter), then the peptides that bind with highest affinity will be present in sufficient quantity to displace the others. Result: A nonrepresentative sampling of the mixture.
TopTips®: Available for samples 1-10, 10-200, and 200-1000 µl.
Their capacity is much greater than that of NuTips or other microscale SPE cartridges. This permits the entire sample to be bound and cleaned up, a safe way to avoid selective loss of components. The following figure shows the removal of 1-2 M urea from a tryptic digest with a HILIC TopTip.
HILIC NuTips and TopTips can also remove detergents from protein and peptide samples.
SPE cartridges: These are for samples up to 5 ml or larger in size. Available in closed cartridges with luer fittings or in packed syringe barrels
(SPE) Cartridges Conditioning:
Attach a syringe to the cartridge and flush with several ml of a solution that promotes retention in the mode employed with the cartridge (see back).
Load thesample solution into the syringe and force it through the cartridge. Flush out non-adsorbed solutes with several ml of the solution used to flush the new cartridge. To recover adsorbed solutes, flush the cartridge with 1-2 ml of a strong eluting solution (see back).Cartridge reuse:In some cases, cartridges can be reused; for example, by flushing out salts with water and then reconditioning with a solution which promotes retention in the mode used. However, the feasibility of reuse must be determined on a case-by-case basis.
Material & ModeLoading & Flushing SolutionEluting SolutionPolyHYDROXYETHYL A™ Hydrophilic Interaction (HILIC)90% ACN + 10 water w/ 5 mM TEAP or ammonium formate, pH 3.020 mM TEAP or ammonium formate, pH 3.0 NO ACN. With unusually hydrophilic solutes, more salts may be necessary.PolyETHYL A™, METHYL A™, & PROPYL A™ Hydrophobic Interaction (HIC)2M ammonium sulfate + 20mM K-PO4, pH 7.0Step to 1M NH4-sulfate + 20 mM K-PO4to elute the weakly hydrophobic solutes; additional step to 20 mM K-PO4to elute to strongly hydrophobic solutesPolySULFOETHYL A™ Cation-Exchange (SCX)5 mM K-PO4, w/ 20% ACN, pH 2.85 mM K-PO4+ 0.5 M KCl, pH 2.8 w/ 20% ACN. To fractionate by basicity, use steps with intermediate levels of KCl.PolyCAT A™ Cation-Exchange (WCX)5 mM K-PO4, w/ 20% ACN, pH 5-65 mM K-PO4+ 0.5 M KCl, pH 5-6 w/ 20% ACN. To fractionate by bascity, use steps with intermediate levels of KCl.PolyWAX LP™ Anion Exchange (WAX)5 mM K-PO4, w/ 20% ACN, pH 75 mM K-PO4+ 0.5 M KCl, pH 7 w/ 20% ACN. To fractionate by bascity, use steps with intermediate levels of KCl.SDS Removal0.1% TFA, or 5 mM K-PO4, pH 2.80.5 M KCl to remove bound SDS prior to reequilibration for r
NuTip® and TopTip® are trademarks of Glygen Corp.
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