LC column, HPLC column, HILIC column, mixed mode column, halo column, gpc column, zorbax column, luna column
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PolyCAT A - for Cation-Exchange of Proteins. PolyCAT A™ is made through a unique process for attaching poly(aspartic acid) covalently to silica. Proteins elute from this polypeptide coating in sharp peaks with little tailing. Binding capacity and recovery are high as well. Use PolyCAT A™ for: 1) Protein variants involving deamidation, PEGylation, position of attachment, desialylation, etc. It is widely used by the biotechnology industry. 2) Hemoglobin variant analysis by clinical chemistry labs. 3) Proteins with pI above 6.0 (5.0 in special cases). 4) Monoclonal antibodies. 5) Histones. Since PolyCAT A™ is a weak cation-exchange (WCX) material, it is used at pH values above 4. A gradient to unbuffered acetic acid will uncharge PolyCAT A™, permitting the elution of proteins in a volatile solvent. Peptides can be run on PolyCAT A™ if they contain at least two excess positive charges above pH 4. More weakly basic peptides, such as tryptic fragments, are not reliably retained and should be run on PolySULFOETHYL A™ at pH = 2.7 - 3.0. For proteins larger than 20 KDa, we recommend the use of pore diameters of at least 1000 Å for optimal selectivity and efficiency. Our 3 µm material with 1000 Å or 1500 Å pores is the finest cation-exchanger available for protein separations.
PolySULFOETHYL A - for Cation-Exchange of Peptides. This strong cation-exchange (SCX) material was developed specifically for HPLC of peptides. At pH 2.7 - 3.0, peptides lose their (-) charges and have net (+) charge. Thus, PolySULFOETHYL A™ is a general-purpose alternative to reverse phase (RPC), fractionating peptides by differences in charge rather than polarity. Compared to other SCX materials based on sulfopropyl- (SP-) groups, PolySULFOETHYL A™ is unusually hydrophilic. This minimizes hydrophobic interactions with peptides, with high recovery and less peak tailing. Capacity is also high, permitting better retention and fractionation of the weakly basic peptides from tryptic digests. The capacity of an ion-exchange material like this is 4x that of a comparable RPC column. Therefore, ion-exchange should be the initial step of a multi-step purification. Standard material for peptide applications is 300 Å, in either 3 µm or 5 µm. The 200 Å material has about 25% greater capacity and is preferred for phosphopeptide isolation and fractionation of iTRAQ® reaction mixtures. For proteins, use 1000 Å material or the 3 µm, 1500 Å material.
PolyWAX LP - for Anion-Exchange of Proteins and Nucleic Acids. 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. 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.
PolyHYDROXYETHYL A - for Hydrophilic Interaction and Size Exclusion HPLC. This neutral, polar material was developed specifically for Hydrophilic Interaction LIquid Chromatography (HILIC). HILIC is a variant of normal phase chromatography that is performed with a polar stationary phase and a partially aqueous mobile phase. This permits normal phase separations of peptides, nucleic acids, carbohydrates, some proteins, and polar solutes in general. In comparison with other materials used for HILIC, PolyHYDROXYETHYL A™ affords sharper peaks and better selectivity and recovery. Its great polarity means less organic solvent is needed to get retention. In the absence of organic solvent, PolyHYDROXYETHYL A™ functions in the Size Exclusion Chromatography (SEC) mode. For proteins and peptides, use 200 Å or 300 Å material. For polar small solutes, try our premium 3 µm, 100 Å material.
Applications
1. Electrostatic Repulsion - Hydrophilic Interaction
Chromatography (ERLIC)
5. Ion Exchange of Proteins with Organic Solvents
8. HILIC Separation
9. Size Exclusion Chromatography (SEC)