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
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
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: