Column

High-Level Protein Expression, One-Column Purification, and FLAG Epitope Tagging in E. coli

Epitope Tagging in E. coli Katherine Felts Denise Wyborski John Bauer PeterVaillancourt Stratagene LIC cloning kit...

Ni-NTA Superflow Columns - automated large-scale purification of 6xHis-tagged proteins

Ni-NTA Superflow Columns enable up to 24 automated large-scale 6xHis-tagged protein preparations to be performed in parallel on the BioRobot 3000 workstation. Using cleared lysates from bacterial cultures as starting material, up to 15 mg protein per column can be purified under native or denaturing conditi...

Purification of Insulin on UNO S1 Column, Rev A

Insulin is a peptide hormone which stimulates anabolic reactions for carbohydrates, proteins, and fats resulting in lowered blood glucose. It is used therapeutically for treatment of diabetes mellitus and was the first recombinant protein on the market. Although insulin is well characterized and understood, it is often difficu...

Purification of Phosphoinositide 3-Kinase With UNO Columns

Polyphosphoinositides and their metabolites are important intracellular signals involved in the responses to a number of hormones and growth factors.The discovery of a phosphatidylinositol kinase that phosphorylates PI at the 3 position of the inositol ring uncovered a new pathway of PI metabolism and potential intracellula...

Purification of Recombinant Flavonone 3beta-Hydroxylase Using Bio-Scale CHT5-1 Hydroxyapatite Column

Flavanone 3b-hydroxylase (FHT), a typical 2-oxoglutaratedependent dioxygenase, catalyzes the 3b-hydroxylation of 2S-flavanones to 2R,3R-dihydroflavonols which are intermediates in the biosynthesis of flavanoids, catechins, proanthocyanidins, and anthocyanidins in plants. To understand the complex reaction mechanism,...

Purification of rIL-13 Using an UNO Q1 Column

Interleukin 13 is a recently described cytokine produced by activated T cells. This protein is responsible for both positive and negative responses to regulatory molecules within the immune system. rIL-13 has been shown to modulate macrophage functions as well as play a direct role in regulation of proliferation and differentiation of hema...

Purification of Two Isoforms of Pyruvate Kinase from Nordic Krill Using an UNO Q1 Column, Rev A

Pyruvate kinase (PK, E.C. 2.7.1.40) is one of the key regulatory enzymes of the glycolytic pathway in crustaceans. This enzyme catalyzes the conversion of phosphoenolpyruvate (PEP) to pyruvate and the phosphorylation of ADP to ATP. In addition to the role of PK in aerobic and anaerobic energy metabolism in...

Comparison of PCR Kleen Spin Columns to Traditional Methods for Purification of PCR Products Prior to Sequencing, Rev A

Jolanta Kolodziejek, MSc, and Professor Norbert Nowotny, PhD, Clinical Virology Group, Institute of Virology, University of Veterinary Medicine, Vienna , Veterinaerplatz 1, A 1210 Vienna, Austria The purity of the DNA template employed in sequencing PCR (cycle sequencing) is the most...

New gravity-flow column gives faster and simpler purification of histidine-tagged proteins

A. Bergh, M. Carlsson, A. Karlsson, and H. Lindgren GE Healthcare, Uppsala, Sweden Histidine-tagged proteins* can be quickly and simply purified on His GraviTrap columns prepacked with Ni Sepharose 6 Fast Flow without the need for a pump or purification system. The protein binding capacity (approx. 40 mg/column) is among the hi...

The combination of monolithic columns and modern HPLC/MS/MS instrumentation for high chromatographic throughput and sensitive detection of beta-agonists

Beta-agonists are group A substances of the European guideline 93/257/EWG (revision 98/536/EG) having an anabolic effect and not licensed. Thus, they have to be determined in veterinarian samples like muscle, liver, retina, and urine. The coupling of High Pressure Liquid Chromatography (HPLC) and Electrospray Ionization Tandem Mass Spectrometry (ESI-MS-MS) seems to be the analytical m...

ANALYSIS OF AMINO ACIDS BY REVERSED PHASE CHROMATOGRAPHY WITH PRECOLUMN DERIVATISATION AND UV/VISIBLE DETECTION

Amino Acids, RP-HPLC, Precolumn derivatisation, o-Phthalaldehyde (OPA), 2-Mercaptopropionic acid, Ultraviolet detection. Introduction: Underivatised amino acids do not significantly absorb light in the UV/visible spectrum. In order to increase sensitivity, their analysis normally includes a derivatisation step in whic...

Three Micron TSK-GEL ODS-100V Columns for High Throughput LCMS

It is well known in HPLC that columns packed with smaller particles are more efficient at the expense of higher back pressure compared to columns packed with larger particles. The gain in efficiency is usually traded off by reducing column length, which in turn results in shorter analysis...

Novel Bonding Chemistry Imparts Enhanced Polar Selectivity to TSK-GEL ODS-100V Reversed Phase Columns

While full coverage C18 phases have been responsible for the growth of HPLC as a reliable high efficiency technique, it has become apparent in recent years that such column types are not suited for reliable analysis of highly polar compounds under low organic or aqueous mobile phase conditions...

TSKgel ODS-100Z and TSKgel ODS-100V column series provides a universal solution for the analysis of samples containing acidic, basic, polar & non-polar compounds

Using high purity Type B silica, Tosoh has introduced two new columns with differing surface treatment procedures and carbon content levels to provide unique selectivities for a wide range of sample types. In recent years, many new octad...

Unique Surface Treatment of Novel TSK-GEL ODS Columns Provide Separation of Aqueous and Fat Soluble Vitamins

With the introduction of two new reversed-phase column types, TSKgel ODS-100V and TSKgel ODS-100Z, excellent chromatographic performance and selectivity can be achieved for a wide range of sample types. TSKgel ODS-100Z and TSKgel ODS-100V are based on a spherical, five micron, silic...

Mobile Phase Optimization for the Analysis of an Antibody-Based Fusion Protein and Aggregates using a TSKgel Super SW3000 Size-Exclusion Column

T.J. Higley and Roy Eksteen, Tosoh Bioscience LLC, Montgomeryville, Pennsylvania andScott Lauder, Lexigen Pharmaceuticals, Lexington, Massachusetts During method development, many variables are examined to ensure method robustness. Factors such as elution profile, peak shape, and recovery...

Improve Recovery and Reproducibility with Pre-Treatment of Analytical TSK-GEL G3000SWXL Size Exclusion Columns

During a recent method optimization and investigation at Wyeth BioPharma, pre-treatment with bovine serum albumin improved reproducibility and recovery during the analytical SEC analysis of fractions pulled from a multi-ste...