PolyWAX LP™ Columns
- for Anion-Exchange of Proteins and Nucleic Acids-
1. Isocratic separation of amino acids, peptides, and proteins.
2. Selective isolation and separation of phosphopeptides.
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Most proteins have isoelectric points below 7, and are best purified or analyzed by anion-exchange chromatography. PolyWAX LP™ is a hydrophilic weak anion-exchange (WAX) material developed by PolyLC for HPLC of enzymes and other proteins. Selectivity is excellent, with high or quantitative recovery of applied biological activity. Most anion-exchange materials based on polyethyleneimine (PEI) are prepared with the conventional branched polymer. PolyWAX LP™ is prepared with linear PEI, which confers greater selectivity and recovery. The following example demonstrates the ability of PolyWAX LP™ to separate proteins differing by a single phosphate group.
Anion-exchange is the method of choice for resolution of oligonucleotides and their analogs > 15 bases. It is also much faster and convenient than PAGE gels for purification of the double-stranded DNA products from PCR reactions.
Use PolyWAX LP™ for:
1) Purification of acidic proteins and polypeptides from natural products.
2) Analysis and purification of oligonucleotides and their analogs as well as amplified PCR products.
3) In proteomics, predigest fractionation of intact proteins via mixed-bed ion-exchange.
4) Anion-exchange of small, acidic solutes.
For small solutes, use our 100-Å material. For peptides, use 300 Å. For proteins > 20 KDa, we recommend pore diameters of at least 1000 Å for optimal selectivity and efficiency. Our 3-µm, 1500-Å material affords superior separations of closely-related protein variants.
The example below shows the elution of a mixture of acidic, basic and neutral peptidesTOP: HILIC mode on a PolyHYDROXYETHYL A™ column. The basic peptides are much better retained than the neutral or basic peptides, since basic solutes are the most polar of all.
BOTTOM: ERLIC mode on a PolyWAX LP™ column. Selective repulsion of the basic peptides throws them into the same elution time frame as the other peptides.