نتایج جستجو برای: capillary isoelectric focusing

تعداد نتایج: 141926  

2014
Noemi Kedei Jin-Qiu Chen Michelle A. Herrmann Andrea Telek Paul K. Goldsmith Mark E. Petersen Gary E. Keck Peter M. Blumberg

Protein kinase C (PKC), a validated therapeutic target for cancer chemotherapy, provides a paradigm for assessing structure-activity relations, where ligand binding has multiple consequences for a target. For PKC, ligand binding controls not only PKC activation and multiple phosphorylations but also subcellular localization, affecting subsequent signaling. Using a capillary isoelectric focusing...

Journal: :Analytical chemistry 2010
Bryan R Fonslow Seong A Kang Daniel R Gestaut Beth Graczyk Trisha N Davis David M Sabatini John R Yates

Here we report the use of capillary isoelectric focusing under native conditions for the separation of protein complex isoforms and subcomplexes. Using biologically relevant HIS-tag and FLAG-tag purified protein complexes, we demonstrate the separations of protein complex isoforms of the mammalian target of rapamycin complex (mTORC1 and 2) and the subcomplexes and different phosphorylation stat...

Journal: :Proceedings of the National Academy of Sciences of the United States of America 2006
Roger A O'Neill Arunashree Bhamidipati Xiahui Bi Debabrita Deb-Basu Linda Cahill Jason Ferrante Erik Gentalen Marc Glazer John Gossett Kevin Hacker Celeste Kirby James Knittle Robert Loder Catherine Mastroieni Michael Maclaren Thomas Mills Uyen Nguyen Nineveh Parker Audie Rice David Roach Daniel Suich David Voehringer Karl Voss Jade Yang Tom Yang Peter B Vander Horn

A previously undescribed isoelectric focusing technology allows cell signaling to be quantitatively assessed in <25 cells. High-resolution capillary isoelectric focusing allows isoforms and individual phosphorylation forms to be resolved, often to baseline, in a 400-nl capillary. Key to the method is photochemical capture of the resolved protein forms. Once immobilized, the proteins can be prob...

Journal: :Analytical chemistry 2007
Zhen Liu Andrei P Drabovich Sergey N Krylov Janusz Pawliszyn

A new method called dynamic kinetic capillary isoelectric focusing (DK-CIEF) is presented for the study of protein-DNA interactions. The method is based on CIEF with laser-induced fluorescence-whole column imaging detection in which protein-DNA complexes are separated with spatial resolution while dissociations of the complexes are dynamically monitored using a CCD camera with temporal resoluti...

Journal: :Clinical chemistry 1994
J M Hempe R D Craver

Capillary isoelectric focusing (cIEF) was used to identify and quantify major and minor hemoglobin (Hb) variants. Whole blood (approximately 10 microL required) hemolysate was analyzed with a commercial instrument equipped with a 50 microns (i.d.) x 27 cm coated capillary filled with 20 g/L ampholytes (pH 6-8) in 4 g/L methylcellulose (MC). Cathode and anode solutions were 20 mol/L NaOH and 100...

Journal: :Lab on a chip 2008
Zhen Liu Junjie Ou Razim Samy Tomasz Glawdel Tiemin Huang Carolyn L Ren Janusz Pawliszyn

Simple-structured, well-functioned disposable poly(dimethylsiloxane) (PDMS) microchips were developed for capillary isoelectric focusing with whole column imaging detection (CIEF-WCID). Side-by-side comparison of the developed microchips with well-established commercial capillary cartridges demonstrated that the disposable microchips have comparable performance as well as advantages such as abs...

2011
David A. Michels Oscar Salas-Solano Chantal Felten

level: interMediAte Analyzing charge variants of therapeutic proteins is critical for characterizing and monitoring quality attributes of antibodies. Charge variants include deamidation, formation of N-terminal pyroglutamate, aggregation, isomerization, sialylated glycans, antibody fragmentation, and glycation at the lysine residues. In some cases, such changes affect binding, biological activi...

2015
Mary G. Johlfs Priyatham Gorjala Yasuyo Urasaki Thuc T. Le Ronald R. Fiscus Ferenc Gallyas

Profiling cellular proteome is critical to understanding signal integration during cell fate determination. In this study, the capability of capillary isoelectric focusing (cIEF) immunoassays to detect post-translational modifications (PTM) of protein isoforms is demonstrated. cIEF immunoassays exhibit protein detection sensitivity at up to 5 orders of magnitude higher than traditional methods....

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