coli) or mammalian cells as a fusion protein with neighboring
affinity tag is one of the most popular methods for purification of protein or protein complex.
The topics include mass spectrometry utilizing isotope-coded
affinity tag reagents, the proteome in neurodegenerative diseases, the proteome related to respiratory disease, hepatitis C virus infection and mitochondria proteomics, and the quantitative proteomic analysis of HIV infection.
However, conventional
affinity tag systems include large protein tags such as green-fluorescent proteins (9 ~ > 27kDa), FLAGs with specific antibodies (9 ~ > 150kDa), an d quantum dots (> ~ 20 [Angstrom]), (1,2) and they affect the affinity to bio-molecules.
In the first laboratory period, each pair of students receives a bacterial lysate containing a pair of unknown fluorescent proteins, one of which possesses an
affinity tag. Over the next few weeks, the students use a bevy of common techniques to identify their two unknowns.
For example, the protein samples are mixed with the ICAT reagents, which consist of a cysteine reactive moiety, a hiotin
affinity tag for isolation and purification, and a linker region consisting of hydrogens (light tag) or deuteriums (heavy tag).
The first five chapters examine the production of proteins for structural biology, specifically discussing the use of a dual
affinity tag for the production of recombinant protein; the cloning, expression, and purification of yeast proteins in a medium-sized structural genomics group, baculovirus expression of integral membrane proteins, and selenomethionine substitution in bacteria and eukaryotic cells.
Complementary analysis of the Mycobacterium tuberculosis proteome by two-dimensional electrophoresis and isotope coded
affinity tag technology.
Oxford GlycoSciences entered a five-year collaboration to develop an industrial-scale proteomics platform using the Institute for Systems Biology's Isotope Coded
Affinity Tag (ICAT) method ...
This novel service is based on a revolutionary new platform, combining
affinity tag binding with inert porous materials, designed especially for biocatalysis application in any organic solvent or aqueous buffer.
The digestion reagents triethylammonium bicarbonate pH 8.5, tris-(2-carboxyethyl)phosphine, and methylmethanethio-sulfate were acquired from Applied Biosystems, as were the isobaric tagging for relative and absolute quantification (iTRAQ) and isotope coded
affinity tag (ICAT) reagents.
Chapters describe studies of protein-protein interactions using near-infrared fluorescence, nuclear magnetic resonance, and mass spectrometry; protein-ligand interactions using mass spectrometry, surface plasmon resonance, nuclear magnetic resonance, and isotope-coded
affinity tag labeling; and structural proteomics using nuclear-magnetic resonance, x-ray crystallography, and electron paramagnetic resonance.
hp vectors are available for intracellular expression, without or with
affinity tag for convenient purification: