Affinity Chromatography - Department of Molecular and Cellular ...
Affinity Chromatography - Department of Molecular and Cellular ...
Affinity Chromatography - Department of Molecular and Cellular ...
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Serine proteases <strong>and</strong> zymogens with an affinity for arginine ........................................... 57<br />
Arginine Sepharose 4B ............................................................................................................................. 57<br />
DNA binding proteins................................................................................................. 59<br />
HiTrap Heparin HP, HiPrep 16/10 Heparin FF, Heparin Sepharose 6 Fast Flow ............................................... 59<br />
Coagulation factors .................................................................................................... 64<br />
HiTrap Heparin HP, HiPrep 16/10 Heparin FF, Heparin Sepharose 6 Fast Flow ............................................... 64<br />
Biotin <strong>and</strong> biotinylated substances .............................................................................. 65<br />
HiTrap Streptavidin HP, Streptavidin Sepharose High Performance ................................................................ 65<br />
Purification or removal <strong>of</strong> fibronectin ........................................................................... 68<br />
Gelatin Sepharose 4B ............................................................................................................................... 68<br />
Purification or removal <strong>of</strong> albumin ............................................................................... 69<br />
HiTrap Blue HP, Blue Sepharose 6 Fast Flow .............................................................................................. 69<br />
NAD + -dependent dehydrogenases <strong>and</strong> ATP-dependent kinases ....................................... 72<br />
5' AMP Sepharose 4B, HiTrap Blue HP, Blue Sepharose 6 Fast Flow ............................................................. 72<br />
NADP + -dependent dehydrogenases <strong>and</strong> other enzymes with affinity for NADP + ............... 74<br />
2'5' ADP Sepharose 4B, Red Sepharose CL-6B ........................................................................................... 74<br />
Glycoproteins or polysaccharides ................................................................................. 79<br />
Con A Sepharose 4B, Lentil Lectin Sepharose 4B, Agarose Wheat Germ Lectin ............................................... 79<br />
Con A for binding <strong>of</strong> branched mannoses, carbohydrates with terminal mannose<br />
or glucose (aMan > aGlc > GlcNAc) .......................................................................................................... 79<br />
Lentil lectin for binding <strong>of</strong> branched mannoses with fucose linked a(1,6) to<br />
the N-acetyl-glucosamine, (aMan > aGlc > GlcNAc) N-acetylglucosamine<br />
binding lectins ......................................................................................................................................... 82<br />
Wheat germ lectin for binding <strong>of</strong> chitobiose core <strong>of</strong> N-linked oligosaccharides,<br />
[GlcNAc(b1,4GlcNAc) 1-2<br />
> b GlcNAc] ......................................................................................................... 83<br />
Calmodulin binding proteins: ATPases, adenylate cyclases,<br />
protein kinases, phosphodiesterases, neurotransmitters ................................................. 85<br />
Calmodulin Sepharose 4B ......................................................................................................................... 85<br />
Proteins <strong>and</strong> peptides with exposed amino acids: His, Cys, Trp,<br />
<strong>and</strong>/or with affinity for metal ions (also known as IMAC,<br />
immobilized metal chelate affinity chromatography) ...................................................... 87<br />
HiTrap Chelating HP, Chelating Sepharose Fast Flow,<br />
His MicroSpin Purification Module, HisTrap Kit ........................................................................................... 87<br />
Thiol-containing substances (purification by covalent chromatography) ........................... 91<br />
Activated Thiol Sepharose 4B, Thiopropyl Sepharose 6B .............................................................................. 91<br />
Chapter 4<br />
Components <strong>of</strong> an affinity medium........................................................................... 96<br />
The matrix ............................................................................................................................................... 96<br />
The lig<strong>and</strong> ............................................................................................................................................... 97<br />
Spacer arms ............................................................................................................................................ 98<br />
Lig<strong>and</strong> coupling ....................................................................................................................................... 99<br />
Lig<strong>and</strong> specificity ..................................................................................................................................... 99<br />
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