Sepax Proteomix Brochure - Winlab.com.au

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Technical SpecificationsProducts Pore Size Particle Size (µm) Dynamic binding capacity pH rangeProteomix SCX-NP1.7, NP3, NP5 & NP10 Non-porous 1.7, 3, 5, 10 ~60, 54, 38, and 20 mg/mL 2-12Proteomix SCX-POR10 500 Å 10 ~35 mg/mL 2-12Proteomix WCX-NP1.7, NP3, NP5 & NP10 Non-porous 1.7, 3, 5, 10 ~25, 19, 15 and 10 mg/mL 2-12Proteomix WCX-POR10 500 Å 10 ~22 mg/mL 2-12Proteomix SAX-NP1.7, NP3, NP5 & NP10 Non-porous 1.7, 3, 5, 10 ~43, 35, 28, and 17 mg/mL 2-12Proteomix SAX-POR10 500 Å 10 ~25 mg/mL 2-12Proteomix WAX-NP1.7, NP3, NP5 & NP10 Non-porous 1.7, 3, 5, 10 ~35, 26, 18, and 12 mg/mL 2-12Proteomix WAX-NOR10 500 Å 10 ~20 mg/mL 2-12ApplicationsSeparation and AnalysisProteinsCell lysatesNucleic acidsSeparation of Horse Serum, a Protein MixtureFigure 12 shows the separation of a horse serum by usingProteomix SAX-NP5 and a commercial porous SAX column.Non-porous Proteomix SAX-NP5 has much high resolutionthan the porous SAX column.Fig. 12. The separation of a horse serum (20 µL, 2x diluted) fromBioWhittaker, a Cambrex company (Walkersville, MD).(Courtesy of Miyako Kawakatsu, M&S Instruments, Inc.)PeptidesCarbohydrates and glycansNucleotidesNanoparticlesNanotubesProteomix ion-exchange phases have wide applications forseparation, identification and purification of proteins, proteinvariants, peptide fragments, phosphorylated, sialylated, andother derivatized proteins. They are excellent for monitoringenzymatic reactions and protein-protein interactions. Withextremely high separation efficiency, Proteomix phases arewell designed for proteomics applications and 2D or multidimensionalseparations. One example is the highresolution of cell lysates by Proteomix SAX-NP phases.Proteomix SAX and WAX phases are suitable for separationof oligonucleotides. Porous Proteomix ion-exchange phasesprovide high resolution and high capacity separation forsurface charged nanoparticles, nanorods and nanotubes.Porous SAX Column X(5 µm, 4.6x150 mm)Proteomix SAX-NP5(5 µm, 4.6x150 mm)Mobile phase: (A) 25 mM Tris acetate, pH 7.3(B) A + 0.8 M sodium acetateGradient: 0-50%-70%B (0-25-28 min)Flow rate: 0.5 mL/min for SAX-NP51.0 mL/min for Porous SAXDetection: UV 280 nmwww.sepax-tech.com 7 1-877-SEPAX-US
Separation of Carbohydrates and GlycansFigure 13 shows the separation of glucose oligomers andcorn starch hydrolysate. ELSD is utilized to detectcarbohydrate molecules. For ELSD detection, bleeding is abig problem for most columns, especially silica based aminocolumns. However, Proteomix SAX columns not only solvedthe bleeding problem, but also achieved excellentseparation efficiency and resolution. Figure 14 is anotherexample of high resolution separation of glycans and theirisomers by non-porous Proteomix SAX column.Fig. 13. Separation of glucose oligomers and corn starchhydrolysate by Proteomix SAX-NP5 phase. (Courtesy of MiyakoKawakatsu, M&S Instruments, Inc.)Starch hydrolysate (5 µL)Glucose oligomer (G1-G6)(5 µL)B, 0.5% acetic acid in ACNGradient: 0-100%B (60 min)Flow rate: 0.3 mL/minDetection: Fluorescence Ex/Em=360/425nmSample: G0F: asialo, agalacto, core-fucosylated biantennaryglycan; G1F: asialo, mono-galacto, core-fucosylated biantennaryglycan; G2F: asialo, di-galacto, core-fucosylated biantennaryglycan; SA1F: mono-sialylated, galactosylated, core-fucosylatedbiantennary glycan; SA2F: di-sialylated, galactosylated, corefucolylatedbiantennary glycan; NGNA: N-glycolylneuraminicacid.Separation of Peptides in Volatile BufferThe peptides with different hydrophobicity can be resolvedby reversed phases, such as C18. However, peptides withdifferent charges may not be well separated on reversedphase mode if their hydrophobic properties are close. As analternative method, ion exchange chromatography isrecommended. With the net charge from +1 to +4, fourpeptides (C1, C2, C3 and C4 with their sequences listed inFigure 15) were well separated by Proteomix SCX-NPphase in a volatile buffer of ammonium acetate andacetonitrile, which makes it compatible for LC/MSapplication.Fig. 15. Cation exchange separation of 4 peptides C1, C2, C3 andC4 (0.1 mg/mL). (Courtesy of Miyako Kawakatsu, M&SInstruments, Inc.)C1C3Column: Proteomix SAX-NP5 (5 µm, 4.6x150 mm)Mobile phase: A, 0.05% (25% NH 4 OH) in CH 3 CNB, 0.05% (25% NH 4 OH) in H 2 OGradient: 15-80%B (20 min)Flow rate: 0.7 mL/minSample: Corn starch hydrolysateTemperature: 35 o CDetection: ELSDC2C4SofTA@280nmFig. 14. Separation of 2-AA (anthranilic acid) labeled N-linkedoligosaccharides profiling of an IgG1 sample.LU4003002001000G0FG1FIsomer of G1FG2FIsomers of SA1F (NGNA)SA2F(NGNA)20 30 40 50 60minColumn: Proteomix SAX-NP5 (5 µm, 4.6x150 mm)Mobile phase: A, 2.5% (v/v) acetic acid, 0.5% TEA in H 2 OColumn: Proteomix SCX-NP3 (3 µm, 4.6x50 mm)Mobile phase: A, 5 mM CH 3 COONH 4B, 500 mM CH 3 COONH 4 :CH 3 CN=4:1(v/v)Gradient: 0-50%B (20 min)Flow rate: 0.6 mL/minTemperature: 25 o CDetection: ELSDInjection: 5 µL (0.1 mg/mL)Sample: Peptides C1, C2, C3 and C4Peptide Sequence Net ChargeC1 Ac-Gly-Gly-Gly-Leu-Gly-Gly-Ala-Gly-Gly-Leu-Lys-amide +1C2C3C4Ac-Lys-Tyr-Gly-Leu-Gly-Gly-Ala-Gly-Gly-Leu-Lys-amide +2Ac-Gly-Gly-Ala-Leu-Lys-Ala-Leu-Lys-Gly-Leu-Lys-amide +3Ac-Lys-Tyr-Ala-Leu-Lys-Ala-Leu-Lys-Gly-Leu-Lys-amide +4minwww.sepax-tech.com 8 1-877-SEPAX-US
Page 3 and 4: Proteomix ® Ion-exchange PhasesGen
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Page 11 and 12: Separation and Analysis of Cell Lys
Page 13 and 14: PhaseID x Length((mm)P/N Price Phas
Page 15: Proteomix WCX-POR ColumnsPhaseID x

Sepax Proteomix Brochure - Winlab.com.au

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