Bind and covalently immobilize antibodies to create a reusable affinity purification column with high binding capacity with Protein A or G agarose columns and DSS crosslinker.
Supplier: Thermo Scientific™ 44893
Thermo Scientific™ Pierce IgG Plus Orientation Kits bind and crosslink antibody to Protein A or Protein G agarose resin, thereby creating a reusable affinity column for purification of antigens.
These IgG orientation kits use a crosslinker to covalently attach antibody that is bound to Protein A or Protein G agarose resin, thereby creating a reusable affinity column for purification of antigens. The Plus Orientation Kits provide for more efficient and stable antibody immobilization than the traditional orientation method because they use the NHS-ester crosslinker DSS (disuccinimidyl suberate) rather than the imidoester crosslinker DMP (dimethyl pimelimidate). Several milligrams of IgG can be bound and crosslinked to each 2mL column, afterwhich the prepared affinity column can be used multiple times for antigen purification using typical binding and elution conditions.The immobilization method is called orientation because antibodies are first captured to the affinity resin through their Fc regions, which are the target binding sites of Protein A and Protein G ligands. Thus, the antibodies are bound with their antigen-binding sites oriented outward. DSS crosslinker reacts with primary amines (side chain of lysine residues) of the Protein A/G and affinity-bound antibody, thereby covalently and permanently attaching the proteins together.
Highlights:
Contaminant-free antigen purification – enables antigen purification without contamination (co-elution) of antibody
Covalent, leak-resistant antibody immobilization – high-capacity binding of IgG to Protein A or Protein G agarose, followed by efficient crosslinking with DSS crosslinker
High-quality agarose resin – crosslinked 6% beaded agarose with recombinant Protein A or Protein G
Gentle binding and crosslinking conditions – coupling antibody in standard PBS
Oriented immobilization – binds and crosslinks IgG with the antigen-binding sites directed outward, able to capture passing antigen in the mobile phase without steric hindrance
Complete kits – include all the reagents needed to immobilize the antibody
Better than DMP – less antibody leaching than traditional orientation method with dimethyl pimelimidate crosslinker
Includes:
Protein A or G columns, disuccinimidyl suberate (DSS), PBS pack, blocking buffer, binding/wash buffer, elution buffer, and column accessories
| 2 mL | |
| Pierce™ | |
| IgG Plus Orientation Kit | |
| 2 to 10 mg of Rabbit IgG per Column | |
| Affinity, Agarose Resin |
| Sufficient For: Immobilizing 2 to 10 mg of rabbit IgG per column • Recombinant Protein A Columns, 2 mL, 2 columns • Disuccinimidyl Suberate (DSS), 2 x 13 mg • PBS Pack (makes 500 mL), 1 pack • Blocking Buffer, 6 mL • IgG Elution Buffer, 15 mL • Antibody Binding/Wash Buffer, 120 mL • Column Accessories Store at 4°C. |
|
| 2 Columns | |
| Protein A | |
| Spin Column |
For Research Use Only. Not for use in diagnostic procedures.
