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Performing a Purification of IgG Antibodies with MabTrap® Kit

MAbTrap® Kit contains one HiTrap® Protein G HP 1 ml column, binding, elution, and neutralization buffers, a syringe with fittings, and an optimized purification protocol (Figure 3.12). The kit contains sufficient material for up to 20 purifications of monoclonal or polyclonal IgG from serum, cell cultur...

Polishing of MAbs Using Capto Adhere ImpRes in Bind/Elute Mode

In these studies, the binding capacity for MAbs and the efficiency in the clearance of impurities using Capto adhere ImpRes in bind/elute mode was evaluated. The studies present results from optimization of the loading conditions using the DoE approach. The effects of buffer, pH, conductivity, and s...

Optimizing Purification of Histidine-Tagged Proteins

The presence of surface-exposed histidine residues or other complex-forming amino acids can lead to unwanted binding of untagged host cell proteins to purification media. These untagged proteins may elute with the target protein. The binding affinity of these contaminants is often lower than that of t...

Performing a Purification of IgG Antibodies with HiTrap® MabSelect™ and HiTrap® MabSelect™ Xtra

This section describes a general procedure for purification of MAbs using HiTrap® MabSelect™ and HiTrap® MabSelect™ Xtra prepacked columns. Figure 3.24 shows capture of mouse monoclonal IgG2a by AC using HiTrap® MabSelect™ followed by an SEC polishing step. The purified MAb is seen in lane 4 of the SD...

Making Immunospecific AC Media with Custom Ligands

If an AC medium is not available, a ligand (such as a pure antigen or an antibody) can be covalently coupled to a suitable matrix to create an immunospecific affinity medium for purification. Although this process requires careful development and optimization, it is often worthwhile, for example when a...

Simple Purification Using a GSTrap Column with ÄKTAprime™

ÄKTAprime, in combination with pre-installed method templates for purifications and prepacked columns, is designed to perform the most common protein purification steps with the touch of one button. It provides significant advantages in speed, capacity, and fraction selection compared with manual pu...

Performing a purification and on-column refolding of an insoluble histidine-tagged protein from a 100 ml E. coli culture using HisTrap FF 1 ml with ÄKTAprime plus

This procedure uses a HisTrap FF 1 ml column but can also be used with a HisTrap HP 1 ml or a HisTrap FF crude 1 ml column. The procedure uses ÄKTAprime plus but can also be run on other ÄKTA systems. Buffers Resuspension buffer 20 mM Tris-HCl, pH 8.0 Isolation buffer 2 M urea, 20 mM Tris-HCl, 0.5 M...

Mesoporous Oxides and Their Applications to Hydrogen Storage

Related ontent • Deposition Grade Silanes for Sol-Gel Processes • • Download PDF • 1Lawrence Berkeley National Laboratory 1 Cyclotron Road, Berkeley, CA 94720 2H2 Technology Consulting LLC 33902 Juliet Circle Fremont, CA 94555 *E-mail: ajhunt@lbl.gov, kgross@h2techconsulting.com, ssmao@newton.berkel...

Chemoselective Purification Tags

Whilst RP-HPLC is an extremely powerful tool for the purification of small to medium sized peptides, for long peptides the technique lacks the resolution necessary to separate the target molecule from the melange of closely related deletion and truncation products that arise during synthesis. In add...

Desalting and Buffer Exchange for Affinity Chromatography of Antibodies

Desalting at laboratory scale is a well-proven, simple, and fast method that will rapidly remove low molecular weight contaminants at the same time as transferring the sample into the desired buffer in a single step. We offer a range of prepacked chromatography columns and 96-well filter plates that ...

Purification Using GSTrap HP, GSTrap FF, and GSTrap 4B columns

Related Articles • Affinity Chromatography in a Purification Strategy (CIPP) • Characteristics of Dextrin Sepharose® High Performance Products • Characteristics of Glutathione Sepharose ® Products • Characteristics of Ni Sepharose®, Ni Sepharose® excel, TALON® Superflow™, and Uncharged IMAC Sepharose®...

Western Blotting Sample Preparation

Symbols This symbol indicates general advice on how to improve procedures or recommends measures to take in specific situations ** indicates a third party trademark The importance of good sample preparation cannot be stressed too highly. By understanding the nature of your starting sample and having...

Cleavage and Purification of GST-Tagged Protein Bound to GSTrap

Recommended buffers Binding buffer: PBS: 140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.3 For PreScission Protease cleavage: Elution buffer: Cleavage buffer: PreScission Protease 50 mM Tris-HCl, 10 mM reduced glutathione, pH 8.0 50 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, pH 7.0 F...

Purification from unclarified cell lysate using HisTrap FF crude

Related Articles • Affinity Chromatography in a Purification Strategy (CIPP) • Characteristics of Dextrin Sepharose® High Performance Products • Characteristics of Glutathione Sepharose ® Products • Characteristics of Ni Sepharose®, Ni Sepharose® excel, TALON® Superflow™, and Uncharged IMAC Sepharose®...

Removal of GST tag by enzymatic cleavage

Related Articles • Affinity Chromatography in a Purification Strategy (CIPP) • Characteristics of Dextrin Sepharose® High Performance Products • Characteristics of Glutathione Sepharose ® Products • Characteristics of Ni Sepharose®, Ni Sepharose® excel, TALON® Superflow™, and Uncharged IMAC Sepharose®...