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Pierce Protein A and Protein G IgG Plus Orientation Kits


Crosslink oriented antibodies to agarose resin to create antigen affinity column.

Pierce IgG Plus Orientation Kits

Thermo Scientific Pierce IgG Plus 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.

These "Plus" 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 2 ml column, afterwhich the prepared affinity column can be used multiple times for antigen purification using typical binding and elution conditions.

Highlights:

  • Contaminant-free antigen purification – provides for 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 DMP crosslinker

Product Details:

Immobilize antibodies by crosslinking them to Protein A or Protein G beads.
Immobilization chemistry of Thermo Scientific IgG Plus Orientation Kits. The crosslinker DSS is used to permanently link antibody that has been bound by immobilized Protein G (or A). Learn more about covalent immobilization methods and about DSS crosslinking chemistry.

References:

  1. Venturi, M., et al. (2000). J. Biol. Chem. 275(7), 4734-4742.

Related Resources:

Review of affinity purification
Review of covalent immobilization methods
Review of immunoprecipitation methods

Related Products:

Pierce Crosslink Immunoprecipitation Kits (small-scale, spin cup version of orientation kits)
Immobilized Protein A (Protein A Agarose)
Immobilized Protein G (Protein G Agarose)
DSS Crosslinker


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
44893 Pierce Protein A IgG Plus Orientation Kit, 2mL
Formulation: Multi-component kit
Sufficient For: Immobilizing 2 to 10mg of rabbit IgG per column

Kit Contents:
Recombinant Protein A Columns, 2mL, 2 columns
Disuccinimidyl Suberate (DSS), 2 x 13mg
PBS Pack (makes 500mL), 1 pack
Blocking Buffer, 6mL
IgG Elution Buffer, 15mL
Antibody Binding/Wash Buffer, 120mL
Column Accessories
2-column kit Product Instructions for product #44893 Pierce Protein A IgG Plus Orientation Kit, 2mL MSDS for product #44893 Pierce Protein A IgG Plus Orientation Kit, 2mL Certificate of Analysis for product #44893 Pierce Protein A IgG Plus Orientation Kit, 2mL $502.00

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44990 Pierce Protein G IgG Plus Orientation Kit, 2mL
Formulation: Multi-component kit
Sufficient For: Immobilizing 1 to 6mg of mouse IgG per column

Kit Contents:
Protein G Columns, 2mL, 2 columns
Disuccinimidyl Suberate (DSS), 2 x 13mg
PBS Pack (makes 500mL), 1 pack
Blocking Buffer, 6mL
IgG Elution Buffer, 15mL
Antibody Binding/Wash Buffer, 120mL
Column Accessories
2-column kit Product Instructions for product #44990 Pierce Protein G IgG Plus Orientation Kit, 2mL MSDS for product #44990 Pierce Protein G IgG Plus Orientation Kit, 2mL Certificate of Analysis for product #44990 Pierce Protein G IgG Plus Orientation Kit, 2mL $628.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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